Predicting mobility using limited data during early stages of a pandemic.

The COVID-19 pandemic has changed consumer behavior substantially. In this study, we explore the drivers of consumer mobility in several metropolitan areas in the United States under the perceived risks of COVID-19. We capture multiple dimensions of perceived risk using local and national cases and death counts of COVID-19, along with real-time Google Trends data for personal protective equipment (PPE). While Google Trends data are popular inputs in many studies, the risk of multicollinearity escalates with the addition of more relevant terms. Therefore, multicollinearity-alleviating methods are needed to appropriately leverage information provided by Google Trends data. We develop and utilize a novel optimization scheme to induce linear models containing strictly significant covariates and minimal multicollinearity. We find that there are a variety of unique factors that drive mobility in different geographic locations, as well as several factors that are common to all locations.

Phosphonium Polymethacrylates for Short Interfering RNA Delivery: Effect of Polymer and RNA Structural Parameters on Polyplex Assembly and Gene Knockdown.

Synthetic polymers containing quaternary phosphonium salts are an emerging class of materials for the delivery of oligo/polynucleotides. In this work, cationic phosphonium salt-containing polymethacrylates and their corresponding ammonium analogues were synthesized by reversible addition-fragmentation chain transfer polymerization. Both the nature of the charged heteroatom (N vs P) and the length of the spacer separating the cationic units along the polymer backbone (oxyethylene vs trioxyethylene) were systematically varied. Polymers efficiently bound short interfering RNA (siRNA) at N(+)/P(-) or P(+)/P(-) ratios of 2 and above. At a 20:1 ratio, small polyplexes (Rh: 4-15 nm) suitable for cellular uptake were formed that displayed low cytotoxicity. While siRNA polyplexes from both ammonium and phosphonium polymers were efficiently internalized by green fluorescent protein (GFP)-expressing 3T3 cells, no knockdown of GFP expression was observed. However, 65% Survivin gene knockdown was observed when siRNA was replaced with novel, multimerized long interfering RNA in HeLa cells, demonstrating the importance of RNA macromolecular architecture on RNA-mediated gene silencing.

Long dsRNA-mediated RNA interference and immunostimulation: a targeted delivery approach using polyethyleneimine based nano-carriers.

RNA oligonucleotides capable of inducing controlled immunostimulation combined with specific oncogene silencing via an RNA interference (RNAi) mechanism provide synergistic inhibition of cancer cell growth. With this concept, we previously designed a potent immunostimulatory long double stranded RNA, referred to as liRNA, capable of executing RNAi mediated specific target gene silencing. In this study, we developed a highly effective liRNA based targeted delivery system to apply in the treatment of glioblastoma multiforme. A stable nanocomplex was fabricated by complexing multimerized liRNA structures with cross-linked branched poly(ethylene imine) (bPEI) via electrostatic interactions. We show clear evidence that the cross-linked bPEI was quite effective in enhancing the cellular uptake of liRNA on U87MG cells. Moreover, the liRNA-PEI nanocomplex provided strong RNAi mediated target gene silencing compared to that of the conventional siRNA-PEI complex. Further, the bPEI modification strategy with specific ligand attachment assisted the uptake of the liRNA-PEI complex on the mouse brain endothelial cell line (b.End3). Such delivery systems combining the beneficial elements of targeted delivery, controlled immunostimulation, and RNAi mediated target silencing have immense potential in anticancer therapy.

Commissioning and dosimetric validation of a novel compensator-based Co-60 IMRT system for evaluating suitability to automated treatment planning.

PURPOSE: A novel compensator-based system has been proposed which delivers intensity-modulated radiation therapy (IMRT) with cobalt-60 beams. This could improve access to advanced radiotherapy in low- and middle-income countries. For this system to be clinically viable and to be adapted into the Radiation Planning Assistant (RPA), being developed to offer automated planning services in low- and middle-income countries, it is necessary to commission and validate it in a commercial treatment planning system (TPS). METHODS: The novel treatment device considered here employs a cobalt-60 source and nine compensators. Each compensator is produced by 3-D printing a thin plastic mold which is then filled on-demand within the machine with reusable 2-mm-diameter spherical tungsten balls. This system was commissioned in the Eclipse TPS and validation tests were conducted with Monte Carlo using Geant4 Application for Tomographic Emission for percentage depth dose, in-plane profiles, penumbra, and IMRT dose validation. And the American Association of Physicists in Medicine Task Group 119 benchmarking testing was performed. Additionally, compensator-based cobalt-60 IMRT plans were created for 46 head-and-neck cancer cases and compared to the linac-based volumetric modulated arc therapy (VMAT) plans used clinically, then dosimetric parameters were evaluated. Beam-on time for each field was calculated. In addition, the measurement was also performed in a limited environment and compared with the Monte Carlo simulations. RESULTS: The differences in percent depth doses and in-plane profiles between the Eclipse and Monte Carlo simulations were 0.65% ± 0.41% and 1.02% ± 0.99%, respectively, and the 80%-20% penumbra agreed within 0.46 ± 0.27 mm. For the Task Group 119 validation plans, all treatment planning goals were met and gamma passing rates were >95% (3%/3 mm criteria). In 46 clinical head-and-neck cases, the cobalt-60 compensator-based IMRT plans had planning target volume (PTV) coverages similar to linac-based VMAT plans: all dosimetric values for PTV were within 1.5%. The organs at risk dose parameters were somewhat higher in cobalt-60 compensator-based IMRT plans versus linac-based VMAT plans. The mean dose differences for the spinal cord, brain, and brainstem were 4.43 ± 1.92, 3.39 ± 4.67, and 2.40 ± 3.71 Gy, while those for the rest of the organs were <1 Gy. The average beam-on time per field was 0.42 ± 0.10 min for the 6 MV multi-leaf-collimator plans while those for the cobalt-60 compensator plans were 0.17 ± 0.01 and 0.31 ± 0.01 min at the dose rates of 350 and 175 cGy/min. There was a good agreement between in-plane profiles from measurements and Monte Carlo simulations, which differences are 1.34 ± 1.90% and 0.13 ± 2.16% for two different fields. CONCLUSIONS: A novel compensator-based IMRT system using cobalt-60 beams was commissioned and validated in a commercial TPS. Plan quality with this system was comparable to that of linac-based plans in all test cases with shorter estimated beam-on times. This system enables reliable, high-quality plans with reduced cost and complexity and may have benefits for underserved regions of the world. This system is being integrated into the RPA, a web-based platform for auto-contouring and auto-planning.

Stem cell derived extracellular vesicles for vascular elastic matrix regenerative repair.

Abdominal aortic aneurysms (AAA) are localized expansions of the abdominal aorta that develop due to chronic proteolytic disruption of the structural extracellular matrix (ECM) components (elastin and collagen) within the aorta wall. Major limitations in arresting or reversing AAAs lie in naturally poor and aberrant regeneration and repair of elastic matrix structures in the aorta wall. Bone marrow derived mesenchymal stem cells (BM-MSCs) have emerged as a promising regenerative tool and their therapeutic effects are also known to be effected through their paracrine secretions. Extracellular vesicles (EVs) present in these secretions have emerged as critical cellular component in facilitating many therapeutic benefits of MSCs. EV treatment is thus potentially appealing as a stem cell-inspired cell-free approach to avoid possible phenotypic plasticity of MSCs in vivo. In this study, we investigated the thus far unknown effects of BM-MSC derived EVs on vascular elastic matrix repair in the context of AAA treatment. EVs isolated from BM-MSC source were characterized and their pro-regenerative and their anti-proteolytic effects were evaluated on our established in vitro experimental conditions derived from AAA rat model. Our studies revealed the efficacy of BM-MSC derived EVs in attenuating the proteolytic activity and also in imparting elastic matrix regenerative benefits under aneurysmal environment. Interestingly, compared to cell culture conditioned media (CCM), EVs demonstrated superior regenerative and anti-proteolytic benefits in a proteolytic injury culture model of AAA. From these studies, it appears that EVs derived from BM-MSCs could be beneficial in undertaking a reparative effort in AAA induced degeneration of vascular tissue. Statement of Significance Abdominal aortic aneurysms (AAAs) are localized, rupture-prone expansions of the aorta which result from loss of wall flexibility due to enzymatic breakdown of elastic fibers. There are no established alternatives to surgery, which possess high risk for the mostly elderly patients. Our previous studies have established the elastic regenerative and reparative effect of cell culture secretions derived from adult stem cell source. In this study, we propose to isolate extracellular vesicles (exosomes) from these secretions and evaluate their regenerative benefits in AAA smooth muscle cell culture model. This simple and innovative treatment approach has the potential to arrest or reverse AAA growth to rupture, not possible so far.

Core-shell hybrid nanostructured delivery platforms for advanced RNAi therapeutics.

AIM: Study was aimed at combining the advantages of nonclassical RNAi-triggering oligonucleotides with nanoparticle-based advanced delivery platforms for developing efficient therapeutic systems. MATERIALS & METHODS: We utilized a core-shell hybrid nanostructured platform for effectively delivering nonclassical RNAi triggers, namely long double stranded interfering RNA and tripodal interfering RNA. Core-shell structure was prepared by stably anchoring thiol-modified cationic polymer on the surface of growing crystal gold (Au) seeds, and the resulting particles were further complexed with nonclassical RNAi candidates via electrostatic interactions. RESULTS: Our studies clearly demonstrated that the unique combination of nonclassical RNAi structures with an advanced core-shell hybrid nanostructured platform is an effective module for advanced RNAi-based therapeutic development.

Cyclodextrin-insulin complex encapsulated polymethacrylic acid based nanoparticles for oral insulin delivery.

Present investigation was aimed at developing an oral insulin delivery system based on hydroxypropyl beta cyclodextrin-insulin (HPbetaCD-I) complex encapsulated polymethacrylic acid-chitosan-polyether (polyethylene glycol-polypropylene glycol copolymer) (PMCP) nanoparticles. Nanoparticles were prepared by the free radical polymerization of methacrylic acid in presence of chitosan and polyether in a solvent/surfactant free medium. Dynamic light scattering (DLS) experiment was conducted with particles dispersed in phosphate buffer (pH 7.4) and size distribution curve was observed in the range of 500-800 nm. HPbetaCD was used to prepare non-covalent inclusion complex with insulin and complex was analyzed by Fourier transform infrared (FTIR) and fluorescence spectroscopic studies. HPbetaCD complexed insulin was encapsulated into PMCP nanoparticles by diffusion filling method and their in vitro release profile was evaluated at acidic/alkaline pH. PMCP nanoparticles displayed good insulin encapsulation efficiency and release profile was largely dependent on the pH of the medium. Enzyme linked immunosorbent assay (ELISA) study demonstrated that insulin encapsulated inside the particles was biologically active. Trypsin inhibitory effect of PMCP nanoparticles was evaluated using N-alpha-benzoyl-L-arginine ethyl ester (BAEE) and casein as substrates. Mucoadhesive studies of PMCP nanoparticles were conducted using freshly excised rat intestinal mucosa and the particles were found fairly adhesive. From the preliminary studies, cyclodextrin complexed insulin encapsulated mucoadhesive nanoparticles appear to be a good candidate for oral insulin delivery.

Novel pH responsive polymethacrylic acid-chitosan-polyethylene glycol nanoparticles for oral peptide delivery.

In present study, novel pH sensitive polymethacrylic acid-chitosan-polyethylene glycol (PCP) nanoparticles were prepared under mild aqueous conditions via polyelectrolyte complexation. Free radical polymerization of methacrylic acid (MAA) was carried out in presence of chitosan (CS) and polyethylene glycol (PEG) using a water-soluble initiator and particles were obtained spontaneously during polymerization without using organic solvents or surfactants/steric stabilizers. Dried particles were analyzed by scanning electron microscopy (SEM) and particles dispersed in phosphate buffer (pH 7.0) were visualized under transmission electron microscope (TEM). SEM studies indicated that PCP particles have an aggregated and irregular morphology, however, TEM revealed that these aggregated particles were composed of smaller fragments with size less than 1 micron. Insulin and bovine serum albumin (BSA) as model proteins were incorporated into the nanoparticles by diffusion filling method and their in vitro release characteristics were evaluated at pH 1.2 and 7.4. PCP nanoparticles exhibited good protein encapsulation efficiency and pH responsive release profile was observed under in vitro conditions. Trypsin inhibitory effect of these PCP nanoparticles was studied using casein substrate and these particles displayed lesser inhibitory effect than reference polymer carbopol. Preliminary investigation suggests that these particles can serve as good candidate for oral peptide delivery.

Guanidine modified polyethyleneimine-g-polyethylene glycol nanocarriers for long interfering RNA (liRNA) based advanced anticancer therapy.

Combination therapy involving the synergism between different therapeutic approaches seems to be a promising strategy in anticancer treatment. Immunostimulatory long interfering RNA (liRNA) structures capable of executing specific RNA interference (RNAi) mediated gene silencing tasks seem to be potential candidates for a combination approach. Apart from their therapeutic efficacy, the unique structural format of liRNA candidates facilitates better association with cationic polymers and significantly improves their intra-cellular delivery. In this study, we have developed a biocompatible cationic delivery platform based on low molecular weight branched polyethyleneimine-grafted-polyethylene glycol (bPEI-g-PEG) for advanced liRNA based anticancer therapy. With simple guanidine (GU) modification, the bPEI-g-PEG platform could induce a strong RNAi mediated response in cancer cells, without induction of any obvious toxicity. Moreover, liRNA complexed with GU-bPEI-g-PEG which targets expression of Survivin gene sensitized cancer cells for effective chemotherapy. A combination strategy involving immunostimulatory RNAi mediators with conventional chemotherapeutic drugs seems to be an effective approach in advanced anticancer treatment.

Poly methacrylic acid-alginate semi-IPN microparticles for oral delivery of insulin: a preliminary investigation.

Microparticles of Poly methacrylic acid (P1) and novel semi-interpenetrating network composed of Poly methacrylic acid-alginate (P2) were prepared and their application in oral insulin delivery was evaluated. The microparticles were characterized by scanning electron microscopy (SEM) for morphological studies. Insulin loading onto the microparticles was performed by the diffusion filling method and insulin encapsulated microparticles were subjected to in vitro release study in buffer solution of pH 1.2 and 7.4. The release kinetics at pH 7.4 exhibited sustained release of insulin for more than 5 h in case of PMAA microparticles whereas burst release of insulin (90% of total insulin loaded) within 1 h of study was observed in the case of PMAA-alginate microparticles. At pH 1.2, around 30% of insulin loaded was released from both microparticles within 2 h of study.

Efficient intracellular delivery and multiple-target gene silencing triggered by tripodal RNA based nanoparticles: a promising approach in liver-specific RNAi delivery.

RNA interference (RNAi) triggering oligonucleotides in unconventional structural format can offer advantages over conventional small interfering RNA (siRNA), enhanced cellular delivery and improved target gene silencing. With this concept, we present a well-defined tripodal-interfering RNA (tiRNA) structure that can induce simultaneous silencing of multiple target genes with improved potency. The tiRNA structure, formed by the complementary association of three single-stranded RNA units, was optimized for improved gene silencing efficacy. When combined with cationic polymers such as linear polyethyleneimine (PEI), tiRNA assembled to form a stable nano-structured complex through electrostatic interactions and induced stronger RNAi response over conventional siRNA-PEI complex. In combination with a liver-targeting delivery system, tripodal nucleic acid structure demonstrated enhanced fluorescent accumulation in mouse liver compared to standard duplex nucleic acid format. Tripodal RNA structure complexed with galactose-modified PEI could generate effective RNAi-mediated gene silencing effect on experimental mice models. Our studies demonstrate that optimized tiRNA structural format with appropriate polymeric carriers have immense potential to become an RNAi-based platform suitable for multi-target gene silencing.

Glucosylated polymeric nanoparticles: a sweetened approach against blood compatibility paradox.

Surface functionalization strategies in generating stealth nano-carriers have garnered considerable attention in pharmaceutical research. In this regard, our investigation reports on the preparation and evaluation of glucose decorated poly lactic-co-glycolic acid (PLGA) nanoparticles as blood compatible nanoparticulate drug delivery system, with enhanced cellular uptake. Terminal carboxylic acid groups on PLGA were modified with the amino group of glucosamine and nanoparticles were generated by modified solvent diffusion (nano-precipitation) technique. Detailed in vitro experiments were performed to evaluate the eminence of glucose functionalization over unmodified nanoparticles, in terms of their hemocompatibility and cellular uptake. Glucosylation was confirmed by NMR and FTIR spectroscopy; PLGA and modified particles had average size in the range of 125nm. Glucosylation was an effective strategy in reducing plasma protein adsorption, complement activation and platelet plugging of PLGA nanoparticles. PLGA and their glucose modified versions were quite compatible with the blood cells and were non-cytotoxic. Moreover the uptake behaviour of glucose modified PLGA nanoparticles were enhanced in comparison to standard PLGA nanoparticles as emphasized by the z stacking analysis following confocal imaging. Precisely the stealth properties of glucose modified PLGA nanoparticles (PLGA-Glu), with enhanced cellular internalization, seems to be a safe and efficient system for intravenous drug delivery applications.

Biomimetic mucin modified PLGA nanoparticles for enhanced blood compatibility.

Efforts to develop long circulating polymeric nanoparticles have propelled many strategies in nanoparticle surface modification to bypass immune surveillance and systemic clearance. In this context, our present study reports on the preparation and evaluation of mucin functionalized poly lactic-co-glycolic acid (PLGA) nanoparticles as hemocompatible, cell penetrating nanoparticulate drug delivery system. Amino groups of mucin were conjugated to the terminal carboxylic acid groups on PLGA to be followed by nanoparticle synthesis via standard solvent evaporation technique. Detailed in vitro experiments were performed to illustrate the significance of alternating copolymer structured mucin modified PLGA nanoparticles in terms of enhanced hemocompatibility and cellular uptake. Mucylation proved promising in controlling PLGA nanoparticle- interaction with plasma proteins (opsonins) and blood components via hemolysis, thrombogenecity and complement activation. Besides hemocompatibility, the modified and unmodified nanoparticles were also found to be cytocompatible with L929 and C6 cell lines. The fluorescent and confocal image analysis evaluated the extent of cellular uptake of nanoparticles into C6 cells. Specifically the combination of stealth properties and cellular internalization capacity of mucin modified PLGA nanoparticle (PLGA-Mucin) lead us to propose it as a safe, efficient and multifunctional nanoplatform for disease specific intravenous drug delivery applications as far as in vitro experiments are concerned.

Mucoadhesive hydrogel microparticles based on poly (methacrylic acid-vinyl pyrrolidone)-chitosan for oral drug delivery.

The study was aimed at the evaluation of N-vinyl pyrrolidone (NVP) incorporated polymethacrylic acid-chitosan microparticles for oral drug delivery applications. Poly (methacrylic acid)-chitosan (PMC) and poly(methacrylic acid-vinyl pyrrolidone)-chitosan (PMVC) microparticles were prepared by an ionic-gelation method. Mucoadhesion behaviour of these particles was evaluated by ex-vivo adhesion method using freshly excised rat intestinal tissue. Cytotoxicity and absorption enhancing property of PMC and PMVC particles were evaluated on Caco 2 cell monolayers. Protease enzyme inhibition capability and insulin loading/release properties of these hydrogel particles was evaluated under in vitro experimental conditions. Addition of NVP units enhanced the mucoadhesion behavior of PMC particles on isolated rat intestinal tissue. Both PMC and PMVC particles were found non-toxic on Caco 2 cell monolayers and PMC particles was more effective in improving paracellular transport of fluorescent dextran across Caco 2 cell monolayers as compared to PMVC particles. However, protease inhibition efficacy of PMC particles was not significantly affected with NVP addition. NVP incorporation improved the insulin release properties of PMC microparticles at acidic pH. Hydrophilic modification seems to be an interesting approach in improving mucoadhesion capability of PMC microparticles.

Effect of thiol functionalization on the hemo-compatibility of PLGA nanoparticles.

In this study, an attempt was made to reduce the interaction of poly(D,L-lactic acid/glycolic acid) (PLGA) nanoparticles with the opsonins and phagocytic cells upon functionalization with thiol groups. Terminal carboxylic groups in PLGA were conjugated to the amino group of cysteine and nanoparticles were prepared by solvent evaporation technique. Detailed in vitro investigations were performed on PLGA and cysteine modified PLGA (Cys-PLGA) nanoparticles to asses their blood compatibility. The effect of these nanoparticles on the release of proinflammatory cytokines (IL-1ß, IL-6, and TNF-α) from human macrophage cells were evaluated. Thiolation was confirmed by fourier transform infrared spectroscopy and Ellman's assay; both PLGA and modified nanoparticles had average size in the range of 250 nm. Thiolation was an effective strategy in reducing the protein adsorption, complement activation, and platelet activation of PLGA nanoparticles. PLGA and modified PLGA nanoparticles were compatible with the blood cells and no hemolytic effect was detected. Particles were noncytotoxic on L929 cells and release of proinflammatory cytokines from macrophage cells was rather unaffected with the modification strategy. From these studies, it seems that thiolation of particulate delivery system is an interesting approach in manipulating the blood-particle interactions and appears to be an effective candidate for injectable drug delivery applications.

Surface-functionalized polymethacrylic acid based hydrogel microparticles for oral drug delivery.

Aim of the present work was to develop novel thiol-functionalized hydrogel microparticles based on poly(methacrylic acid)-chitosan-poly(ethylene glycol) (PCP) for oral drug delivery applications. PCP microparticles were prepared by a modified ionic gelation process in aqueous medium. Thiol modification of surface carboxylic acid groups of PCP micro particles was carried out by coupling l-cysteine with a water-soluble carbodiimide. Ellman's method was adopted to quantify the sulfhydryl groups, and dynamic light-scattering technique was used to measure the average particle size. Cytotoxicity of the modified particles was evaluated on Caco 2 cells by MTT assay. Effect of thiol modification on permeability of paracellular marker fluorescence dextran (FD4) was evaluated on Caco 2 cell monolayers and freshly excised rat intestinal tissue with an Ussing chamber set-up. Mucoadhesion experiments were carried out by an ex vivo bioadhesion method with excised rat intestinal tissue. The average size of the PCP microparticles was increased after thiol modification. Thiolated microparticles significantly improved the paracellular permeability of FD4 across Caco 2 cell monolayers, with no sign of toxicity. However, the efficacy of thiolated system remained low when permeation experiments were carried out across excised intestinal membrane. This was attributed to the high adhesion of the thiolated particles on the gut mucosa. Nevertheless, it can be concluded that surface thiolation is an interesting strategy to improve paracellular permeability of hydrophilic macromolecules.

Cyclodextrin complexed insulin encapsulated hydrogel microparticles: An oral delivery system for insulin.

An oral insulin delivery system based on methyl-ß-cyclodextrin (MCD) complexed insulin encapsulated polymethacrylic acid (PMAA) hydrogel microparticles was evaluated in this investigation. Poly(methacrylic acid)-chitosan-polyethylene glycol (PCP) microparticles were prepared by ionic gelation method. The insulin-MCD (IC) complex prepared was characterized by fluorescence spectroscopic and isothermal titration micro-calorimeteric (ITC) methods. MCD complexed insulin was encapsulated onto PCP microparticles by diffusion filling method. Loading and release properties of the complexed insulin from microparticles were evaluated under in vitro conditions. The effect of MCD complexation on the permeability of insulin was studied using Caco 2 cell monolayers and excised intestinal tissue with an Ussing chamber set-up. In vivo experiments were carried on streptozotocin induced diabetic rats to evaluate the efficacy of MCD complexed insulin encapsulated PCP microparticles to deliver insulin by the oral route. IC complex formation was established by fluorescence and ITC investigations. Insulin loading and release properties from the hydrogel matrix was rather unaffected by the MCD complexation. However MCD complexation was effective in enhancing insulin transport across Caco 2 cell monolayers, when applied in combination with the PMAA hydrogel system. Both insulin and MCD complexed insulin encapsulated PCP microparticles were effective in reducing blood glucose level in diabetic animal models. Cyclodextrin complexed insulin encapsulated hydrogel microparticles appear to be an interesting candidate for oral delivery of insulin.

Thiol functionalized polymethacrylic acid-based hydrogel microparticles for oral insulin delivery.

In the present study thiol functionalized polymethacrylic acid-polyethylene glycol-chitosan (PCP)-based hydrogel microparticles were utilized to develop an oral insulin delivery system. Thiol modification was achieved by grafting cysteine to the activated surface carboxyl groups of PCP hydrogels (Cys-PCP). Swelling and insulin loading/release experiments were conducted on these particles. The ability of these particles to inhibit protease enzymes was evaluated under in vitro experimental conditions. Insulin transport experiments were performed on Caco-2 cell monolayers and excised intestinal tissue with an Ussing chamber set-up. Finally, the efficacy of insulin-loaded particles in reducing the blood glucose level in streptozotocin-induced diabetic rats was investigated. Thiolated hydrogel microparticles showed less swelling and had a lower insulin encapsulation efficiency as compared with unmodified PCP particles. PCP and Cys-PCP microparticles were able to inhibit protease enzymes under in vitro conditions. Thiolation was an effective strategy to improve insulin absorption across Caco-2 cell monolayers, however, the effect was reduced in the experiments using excised rat intestinal tissue. Nevertheless, functionalized microparticles were more effective in eliciting a pharmacological response in diabetic animal, as compared with unmodified PCP microparticles. From these studies thiolation of hydrogel microparticles seems to be a promising approach to improve oral delivery of proteins/peptides.