Thermosensitive Chitosan-based Hydrogel Research Articles

Enhanced diabetic foot ulcer treatment with a chitosan-based thermosensitive hydrogel loaded self-assembled multi-functional nanoparticles for antibacterial and angiogenic effects

Inhibiting bacterial growth and promoting angiogenesis are essential for enhancing wound healing in diabetic patients. Excessive oxidative stress at the wound site can also lead to an accumulation of reactive oxygen species. To address these challenges, a smart thermosensitive hydrogel loaded with therapeutic agents was developed. This formulation features self-assembled nanoparticles named CIZ, consisting of chlorogenic acid (CA), indocyanine green (ICG), and zinc ions (Zn2+). These nanoparticles are loaded into a chitosan-β-glycerophosphate hydrogel, named CIZ@G, which enables rapid gel formation under photothermal effects. The hydrogel demonstrates good biocompatibility and effectively releases drugs into diabetic foot ulcers (DFU) wound. Benefiting from the dual actions of CA and zinc ions, the hydrogel exhibits potent antioxidative and anti-inflammatory effects, enhances the expression of vascular endothelial growth factor (VEGF) and Platelet endothelial cell adhesion molecule-1 (CD31), and promotes angiogenesis. Both in vitro and in vivo experiments confirm that CIZ@G can effectively inhibit the growth of Staphylococcus aureus post-laser irradiation and accelerate wound remodeling within 14 days. This approach offers a new strategy for the treatment of diabetic foot ulcers (DFU), potentially transforming patient care in this challenging clinical area.

Thermosensitive chitosan-based hydrogel: A vehicle for overcoming the limitations of nose-to-brain cell therapy

Cell therapy is a promising strategy for treating neurological pathologies but requires invasive methods to bypass the blood–brain barrier restrictions. The nose-to-brain route has been presented as a direct and less invasive alternative to access the brain. The primary limitations of this route are low retention in the olfactory epithelium and poor cell survival in the harsh conditions of the nasal cavity. Thus, using chitosan-based hydrogel as a vehicle is proposed in this work to overcome the limitations of nose-to-brain cell administration. The hydrogelʼs design was driven to achieve gelification in response to body temperature and a mucosa-interacting chemical structure biocompatible with cells. The hydrogel showed a < 30 min gelation time at 37 °C and >95 % biocompatibility with 2D and 3D cultures of mesenchymal stromal cells. Additionally, the viability, stability, and migration capacity of oligodendrocyte precursor cells (OPCs) within the hydrogel were maintained in vitro for up to 72 h. After the intranasal administration of the OPCs-containing hydrogel, histological analysis showed the presence of viable cells in the nasal cavity for up to 72 h post-administration in healthy athymic mice. These results demonstrate the hydrogel's capacity to increase the residence time in the nasal cavity while providing the cells with a favorable environment for their viability. This study presents for the first time the use of thermosensitive hydrogels in nose-to-brain cell therapy, opening the possibility of increasing the delivery efficiency in future approaches in translational medicine. Statement of significanceThis work highlights the potential of biomaterials, specifically hydrogels, in improving the effectiveness of cell therapy administered through the nose. The nose-to-brain route has been suggested as a non-invasive way to directly access the brain. However, delivering stem cells through this route poses a challenge since their viability must be preserved and cells can be swept away by nasal mucus. Earlier attempts at intranasal cell therapy have shown low efficiency, but still hold promise to the future. The hydrogels designed for this study can provide stem cells with a biocompatible environment and adhesion to the nasal atrium, easing the successful migration of viable cells to the brain.

Thermosensitive hydrogel doped with osteoconductive fillers for the treatment of periodontitis periapicalis chronica: from synthesis to clinical trial.

Herein, a chitosan-based thermosensitive hydrogel (CH) containing hydroxyapatite (HAp), poly(lactic acid) (PLDLLA) or their mixture is proposed as an innovative, biomimetic composition with antimicrobial and bone-forming properties for guided bone regeneration. The modified hydrogels were synthesized and characterized to verify their suitability for the treatment of periodontitis periapicalis chronica. Compared to the unmodified hydrogel, both CH_HAp and CH_PLDLLA revealed improved mechanical properties, as evidenced by rotational rheology. FTIR analysis proved that no chemical interplay existed between the components. All the tested samples displayed no cytotoxicity against osteoblast-like cell culture and confirmed antimicrobial features, both crucial from an application perspective. Radiation sterilization dosage was tailored for the tested samples to maintain sterility for a minimum of 8 weeks of storage and limit crosslinking of the samples. Finally, the hydrogel was used in a clinical trial to treat a patient with chronic inflammation of periapical tissues in teeth 26 and 27. The medical procedure proved the safety, nontoxicity, non-allergenicity, and, most importantly, bone-forming properties of the hydrogel formulation. The kinetics of new bone formation was analyzed in-depth using graphical cross-sections of anatomical structures obtained from pre- and post-operative CBCT scans.

Mesenchymal stem cell derived extracellular vesicles loaded thermosensitive chitosan-based hydrogel alleviates allergic rhinitis in mouse model

Allergic rhinitis (AR) has gained an increasing prevalence over the years, a more effective and safe treatment strategy need to be carried out. Mesenchymal stem cell derived extracellular vesicles (MSC-EVs) show great therapeutic potential for AR, however, their low bioavailability through currently available delivery methods inhibits the process of clinical application. In the present study, an injectable thermosensitive chitosan based hydrogel was developed as an intranasal delivery platform for bone marrow-derived MSC-EVs (BMSC-EVs) and BMSC-EVs were proved to be effective in treating AR animal models for the first time. Our results showed that the BMSC-EVs loaded thermosensitive chitosan based hydrogel (BMSC-EVs@CHI hydrogel) could be prepared easily by ion-crosslinking and its gelation properties were suitable for intranasal administration. In vitro studies demonstrated that the BMSC-EVs@CHI hydrogel could achieve sustained release of BMSC-EVs mainly by degradation control and preserve the therapeutic functions of BMSC-EVs. An in vivo experiment revealed that the BMSC-EVs@CHI hydrogel could enhance the efficacy of BMSC-EVs for AR alleviation. This study provides a suitable delivery platform for BMSC-EVs-based AR therapy.

Chitosan-based thermosensitive hydrogel with long-term release of murine nerve growth factor for neurotrophic keratopathy.

Neurotrophic keratopathy is a persistent defect of the corneal epithelium, with or without stromal ulceration, due to corneal nerve deficiency caused by a variety of etiologies. The treatment options for neurotrophic keratopathy are limited. In this study, an ophthalmic solution was constructed from a chitosan-based thermosensitive hydrogel with long-term release of murine nerve growth factor (CTH-mNGF). Its effectiveness was evaluated in corneal denervation (CD) mice and patients with neurotrophic keratopathy. In the preclinical setting, CTH-mNGF was assessed in a murine corneal denervation model. CTH-mNGF was transparent, thermosensitive, and ensured sustained release of mNGF for over 20 hours on the ocular surface, maintaining the local mNGF concentration around 1300 pg/mL in vivo. Corneal denervation mice treated with CTH-mNGF for 10 days showed a significant increase in corneal nerve area and total corneal nerve length compared with non-treated and CTH treated mice. A subsequent clinical trial of CTH-mNGF was conducted in patients with stage 2 or 3 neurotrophic keratopathy. Patients received topical CTH-mNGF twice daily for 8 weeks. Fluorescein sodium images, Schirmer's test, intraocular pressure, Cochet-Bonnet corneal perception test, and best corrected visual acuity were evaluated. In total, six patients (total of seven eyes) diagnosed with neurotrophic keratopathy were enrolled. After 8 weeks of CTH-mNGF treatment, all participants showed a decreased area of corneal epithelial defect, as stained by fluorescence. Overall, six out of seven eyes had fluorescence staining scores < 5. Moreover, best corrected visual acuity, intraocular pressure, Schirmer's test and Cochet-Bonnet corneal perception test results showed no significant improvement. An increase in corneal nerve density was observed by in vivo confocal microscopy after 8 weeks of CTH-mNGF treatment in three out of seven eyes. This study demonstrates that CTH-mNGF is transparent, thermosensitive, and has sustained-release properties. Its effectiveness in healing corneal epithelial defects in all eyes with neurotrophic keratopathy suggests CTH-mNGF has promising application prospects in the treatment of neurotrophic keratopathy, being convenient and cost effective.

Construction of chitosan-based thermosensitive composite hydrogels for recognizing and combined chemo-photodynamic elimination of Gram-negative bacterial infections

Recently, rapid acquisition of bacterial resistance and consequent slow healing of infected wounds threaten human life and health. In this study, chitosan-based hydrogels and nanocomplexes ZnPc(COOH)8:PMB composed of photosensitizer ZnPc(COOH)8 and antibiotic polymyxin B (PMB) were integrated into a thermosensitive antibacterial platform ZnPc(COOH)8:PMB@gel. Interestingly, fluorescence and reactive oxygen species (ROS) of ZnPc(COOH)8:PMB@gel can be triggered by E. coli bacteria at 37 °C, but not by S. aureus bacteria, which gave the potential to simultaneously detect and treat Gram-negative bacteria. The survival rate for a certain amount of E. coli bacteria treated with ZnPc(COOH)8:PMB (ZnPc(COOH)8 2 μM) was decreased by approximately fivefold than that with either ZnPc(COOH)8 or PMB alone, indicating combined antibacterial efficacy. ZnPc(COOH)8:PMB@gel facilitated the complete healing of wounds infected with E. coli bacteria in about seven days, while over 10 % wounds treated with ZnPc(COOH)8 or PMB remained unhealed on the 9th day. ZnPc(COOH)8:PMB resulted in a threefold increase of ZnPc(COOH)8 fluorescence in E. coli bacteria suggesting enhanced uptake of ZnPc(COOH)8 for the intervention of PMB on membrane permeability. The construction principle of the thermosensitive antibacterial platform and the combined antimicrobial strategy can be applied to other photosensitizers and antibiotics for detection and treatment of wound infections.

An injectable multifunctional thermo-sensitive chitosan-based hydrogel for periodontitis therapy

Bacteria are recognized as the driving factors of periodontitis. However, excessive reactive oxygen species (ROS) can harm periodontal tissue while also causing an uncontrolled inflammatory response. Hence, eliminating excessive ROS and blocking ROS-induced abnormal inflammatory response by antioxidants are achieving remarkable results in periodontitis therapy. Moreover, influenced by the deep and irregular periodontal pockets, injectable thermo-sensitive chitosan-based hydrogels have attracted a lot of attention. This study aimed to formulate an antibacterial and antioxidant therapeutic regimen by incorporating antimicrobial peptides (Nal-P-113) and/or antioxidants (polydopamine nanoparticles, PDNPs) into chitosan-based hydrogels. The hydrogel was characterized in vitro and finally examined in rats using the experimental periodontitis model. The release kinetics showed that the hydrogel could stably release Nal-P-113 and PDNPs for up to 13 days. The scavenging activity of the hydrogel against DPPH was about 80 % and the antibacterial ratio against Streptococcus gordonii (S. gordonii), Fusobacterium nucleatum (F. nucleatum) and Porphyromonas gingivalis (P. gingivalis) was about 99 %. Importantly, it was examined that the hydrogel had the ability to prevent periodontal tissue damage. Thus, chitosan-based hydrogels may provide a basis for designing multifunctional local drug delivery biomaterials for the treatment of periodontitis.

Preparation and performance of chitosan/cyclodextrin-g-glutamic acid thermosensitive hydrogel

Chitosan hydrogel had been widely used in cell culture and tissue engineering, but its long gel time, weak mechanical strength, and weak antibacterial performance limited its application. In this study, the cyclodextrin grafted glutamic acid, a water-soluble cyclodextrin derivatives, was prepared by glutamic acid (Glu) and mono-(6-OTs)-β-CD under alkaline conditions and confirmed by FTIR and 1H NMR. Then chitosan (CS) and cyclodextrin grafted glutamic acid (CD-g-Glu) were used to prepare chitosan/cyclodextrin derivatives thermosensitive hydrogel (CS/CD-g-Glu) to improve the properties of chitosan-based thermosensitive hydrogels. The CS/CD-g-Glu hydrogel had temperature sensitivity and stable three-dimensional network structure. In addition, the hydrogel loaded with berberine hydrochloride (BBH) showed optimal sustained-release effect, and the cumulative release rate was 57% in 24 h. More importantly, the CS/CD-g-Glu hydrogel loaded with BBH showed prefect antibacterial activity against E. coli and S. aureus. Therefore, the CS/CD-g-Glu hydrogel has great promising applications in drug delivery, tissue engineering, biotechnological, and biomedical fields.

Controlled Liposome Delivery from Chitosan-Based Thermosensitive Hydrogel for Regenerative Medicine.

This work describes the development of an injectable nanocomposite system based on a chitosan thermosensitive hydrogel combined with liposomes for regenerative medicine applications. Liposomes with good physicochemical properties are prepared and embedded within the chitosan network. The resulting nanocomposite hydrogel is able to provide a controlled release of the content from liposomes, which are able to interact with cells and be internalized. The cellular uptake is enhanced by the presence of a chitosan coating, and cells incubated with liposomes embedded within thermosensitive hydrogels displayed a higher cell uptake compared to cells incubated with liposomes alone. Furthermore, the gelation temperature of the system resulted to be equal to 32.6 °C; thus, the system can be easily injected in the target site to form a hydrogel at physiological temperature. Given the peculiar performance of the selected systems, the resulting thermosensitive hydrogels are a versatile platform and display potential applications as controlled delivery systems of liposomes for tissue regeneration.

Exosomes-loaded thermosensitive hydrogels for corneal epithelium and stroma regeneration

Corneal damage forms scar tissue and manifests as permanent corneal opacity, which is the main cause of visual impairment caused by corneal diseases. To treat these diseases, herein, we developed a novel approach based on the exosome derived from induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) combined with a thermosensitive hydrogel, which reduces scar formation and accelerates the healing process. We found that a thermosensitive chitosan-based hydrogels (CHI hydrogel) sustained-release iPSC-MSC exosomes can effectively promote the repair of damaged corneal epithelium and stromal layer, downregulating mRNA expression coding for the three most enriched collagens (collagen type I alpha 1, collagen type V alpha 1 and collagen type V alpha 2) in corneal stroma and reducing scar formation in vivo. Furthermore, iPSC-MSCs secrete exosomes that contain miR-432-5p, which suppresses translocation-associated membrane protein 2 (TRAM2), a vital modulator of the collagen biosynthesis in the corneal stromal stem cells to avert the deposition of extracellular matrix (ECM). Our findings indicate that iPSC-MSCs secrete miRNA-containing exosomes to promote corneal epithelium and stroma regeneration, and that miR-432-5p can prevent ECM deposition via a mechanism most probably linked to direct repression of its target gene TRAM2. Overall, our exosomes-based thermosensitive CHI hydrogel, is a promising technology for clinical therapy of various corneal diseases.

Rational formulation design of injectable thermosensitive chitosan-based hydrogels for cell encapsulation and delivery

This work reports a rational design of injectable thermosensitive chitosan systems for cell encapsulation and delivery. Using mixtures of two phosphate salts, beta-glycerophosphate and ammonium hydrogen phosphate, we demonstrate that the pH and the osmolarity can be adjusted separately by varying the molar ratios between the salts and the d-glucosamine monomers. We found the existence of a critical temperature above which gelation time decays following a power-law. This gelation kinetics can be finely tuned through the pH and salt-glucosamine ratios. Formulations having physiological pH and osmolarity were produced for chitosan concentrations ranging from 0.4 to 0.9 wt%. They remain liquid for more than 2 h at 20 °C and form a macroporous gel within 2 min at 37 °C. In vitro encapsulation of pre-osteoblastic cells and gingival fibroblasts showed homogeneous cell distribution and good cell viability up to 24 h. Such an approach provides a valuable platform to design thermosensitive cell-laden systems.

Pharmacological Preconditioning Improves the Viability and Proangiogenic Paracrine Function of Hydrogel-Encapsulated Mesenchymal Stromal Cells.

The efficacy of cell therapy is limited by low retention and survival of transplanted cells in the target tissues. In this work, we hypothesize that pharmacological preconditioning with celastrol, a natural potent antioxidant, could improve the viability and functions of mesenchymal stromal cells (MSC) encapsulated within an injectable scaffold. Bone marrow MSCs from rat (rMSC) and human (hMSC) origin were preconditioned for 1 hour with celastrol 1 μM or vehicle (DMSO 0.1% v/v), then encapsulated within a chitosan-based thermosensitive hydrogel. Cell viability was compared by alamarBlue and live/dead assay. Paracrine function was studied first by quantifying the proangiogenic growth factors released, followed by assessing scratched HUVEC culture wound closure velocity and proliferation of HUVEC when cocultured with encapsulated hMSC. In vivo, the proangiogenic activity was studied by evaluating the neovessel density around the subcutaneously injected hydrogel after one week in rats. Preconditioning strongly enhanced the viability of rMSC and hMSC compared to vehicle-treated cells, with 90% and 75% survival versus 36% and 58% survival, respectively, after 7 days in complete media and 80% versus 64% survival for hMSC after 4 days in low serum media (p < 0.05). Celastrol-treated cells increased quantities of proangiogenic cytokines compared to vehicle-pretreated cells, with a significant 3.0-fold and 1.8-fold increase of VEGFa and SDF-1α, respectively (p < 0.05). The enhanced paracrine function of preconditioned MSC was demonstrated by accelerated growth and wound closure velocity of injured HUVEC monolayer (p < 0.05) in vitro. Moreover, celastrol-treated cells, but not vehicle-treated cells, led to a significant increase of neovessel density in the peri-implant region after one week in vivo compared to the control (blank hydrogel). These results suggest that combining cell pretreatment with celastrol and encapsulation in hydrogel could potentiate MSC therapy for many diseases, benefiting particularly ischemic diseases.

New application of an old drug proparacaine in treating epilepsy via liposomal hydrogel formulation

Proparacaine (PPC) is a previously discovered topical anesthetic for ophthalmic optometry and surgery by blocking the central Nav1.3. In this study, we found that proparacaine hydrochloride (PPC-HCl) exerted an acute robust antiepileptic effect in pilocarpine-induced epilepsy mice. More importantly, chronic treatment with PPC-HCl totally terminated spontaneous recurrent seizure occurrence without significant toxicity. Chronic treatment with PPC-HCl did not cause obvious cytotoxicity, neuropsychiatric adverse effects, hepatotoxicity, cardiotoxicity, and even genotoxicity that evaluated by whole genome-scale transcriptomic analyses. Only when in a high dose (50 mg/kg), the QRS interval measured by electrocardiography was slightly prolonged, which was similar to the impact of levetiracetam. Nevertheless, to overcome this potential issue, we adopt a liposome encapsulation strategy that could alleviate cardiotoxicity and prepared a type of hydrogel containing PPC-HCl for sustained release. Implantation of thermosensitive chitosan-based hydrogel containing liposomal PPC-HCl into the subcutaneous tissue exerted immediate and long-lasting remission from spontaneous recurrent seizure in epileptic mice without affecting QRS interval. Therefore, this new liposomal hydrogel formulation of proparacaine could be developed as a transdermal patch for treating epilepsy, avoiding the severe toxicity after chronic treatment with current antiepileptic drugs in clinic.

Development of a dual delivery of levofloxacin and prednisolone acetate via PLGA nanoparticles/ thermosensitive chitosan-based hydrogel for postoperative management: An in-vitro and ex-vivo study

Post-operative endophthalmitis (POE) is one of the most dreadful complications after intraocular surgery. For cataract surgery patients, both commercially available topical 0.5% levofloxacin and 1% prednisolone acetate (PA) ophthalmic solution require at least 3 to 4 times application daily. In this study, we develop a dual drug delivery system composed of the thermosensitive chitosan/gelatin-based hydrogel containing PA and levofloxacin-loaded nanoparticles (LNPs). LNPs with negative surface charge show the monodisperse (polydispersity index ~0.045), nanosize (~154.7 nm) and sphere-like structure. The optimal concentration of LNPs and PA to corneal epithelial cells was 5 μg/mL and 50 μg/mL, respectively. The developed dual drug delivery system (PAgel-LNPs) could gel at 34 °C within 63 s. The osmolarity of PAgel-LNPs was 301.2 ± 1.5 mOsm/L. PAgel-LNPs showed a sustained-release profile for 7 days. Post-treatment of PAgel-LNPs in TNF-α-damaged corneal epithelial cells could decrease the inflammation (inflammatory genes (TNF-α, IL-6, MMP-3 andMMP-9) and IL-6 production) and cell death. In ex-vivo rabbit model of S. aureus keratitis, the anti-inflammation and anti-bacterial property have been demonstrated. These results suggest that thermosensitive PAgel-LNPs may have the potential to use for the prevention of POE.

Chitosan-based thermosensitive hydrogel entrapping calcein for visualizing localized drug delivery

Localized drug delivery (LDD) is one of the unmet needs for the treatment of various localized medical conditions like infections, cancer, pain, etc. Several hydrogel-based modalities are now being developed for drug delivery, tissue regeneration, treating diseases, and postoperative physiological response. Local anesthetics are one of the most widely used medicine in pain management. Unfortunately, there are several drawbacks associated with local anesthetics such as repeated dosing, short duration of action, and dose dumping. However, some drug carriers based on nanoparticles (NPs) have been used to overcome the problems associated with local anesthetics, but these NPs exhibit minimal residence time at the injected site. In the current study, we report a thermosensitive hydrogel based on chitosan and β-glycerophosphate as a cross-linker can be a promising approach for sustained and localized drug delivery drugs. These hydrogels exhibited a phase transition from sol (4 °C) to gel at physiological temperature (37 °C) that can prolong the half-lives by sustained release of the local anesthetics. The hydrogel entrapping calcein (hydrophilic model drug) exhibited sustained release, and by utilizing its fluorescence, its release from the gel was visually monitored in real-time.

Catechol functionalized chitosan/active peptide microsphere hydrogel for skin wound healing

Chitosan-based thermosensitive hydrogels have been widely used in drug delivery and tissue engineering, but their poor bioactivity has limited their further applications. Integral active oyster peptide microspheres (OPM) with an average particle diameter of 3.9 μm were prepared with high encapsulation efficiency (72.8%) and loading capacity (11.9%), exhibiting desirable sustained release effects. Using catechol functionalized chitosan (CS-C) as the polymeric matrix, OPM as the filler, and β-sodium glycerophosphate (β-GP) as a thermal sensitizer, the thermosensitive hydrogel CS-C/OPM/β-GP was prepared. Besides, the application of the hydrogel on wound healing was studied, and its biosafety was evaluated. The results of cell migration in vitro showed that the cell migration rate of CS-C/OPM/β-GP reached 97.47 ± 5.41% within 48 h, indicating that the hydrogel accelerated the migration of L929 cells. As demonstrated in the mouse skin wound experiment, CS-C/OPM/β-GP hydrogel not only inhibited the aggregation of diversified inflammatory cells and accelerated the generation of collagen fibers and new blood vessels of the wound, but also enhanced the synthesis of total protein (TP) in granulation tissue, and up-regulated the expression of Ki-67 and VEGF in the injury, thereby achieving fast wound healing. Safety evaluation results showed that CS-C/OPM/β-GP hydrogel was not cytotoxic to L929 cells, and the hemolysis ratio was less than 5% within 1 mg/mL. In conclusion, CS-C/OPM/β-GP hydrogel is expected as a promising medical dressing for wound healing.

Thermosensitive chitosan-based hydrogels supporting motor neuron-like NSC-34 cell differentiation.

Motor neuron diseases are neurodegenerative diseases that predominantly affect the neuromuscular system. To date, there are no valid therapeutic treatments for such diseases, and the classical experimental models fail in faithfully reproducing the pathological mechanisms behind them. In this regard, the use of three-dimensional (3D) culture systems, which more closely reproduce the native in vivo environment, can be a promising approach. Hydrogel-based systems are among the most used materials to reproduce the extracellular matrix, featuring an intrinsic similarity with its physiological characteristics. In this study, we developed a thermosensitive chitosan-based hydrogel combined with β-glycerophosphate (βGP) and sodium hydrogen carbonate (SHC), which give the system optimal mechanical properties and injectability, inducing the hydrogel sol-gel transition at 37 °C. An ad hoc protocol for the preparation of the hydrogel was established in order to obtain a highly homogeneous system, leading to reproducible physicochemical characteristics and easy cell encapsulation. All formulations supported the viability of a neuroblastoma/spinal cord hybrid cell line (NSC-34) beyond two weeks of culture and enabled cell differentiation towards a motor neuron-like morphology, characterized by the presence of extended neurites. Based on our results, these hydrogels represent excellent candidates for establishing 3D in vitro models of motor neuron diseases.

Fabrication and Characterization of Chitosan Based Injectable Thermosensitive Hydrogels Containing Silica/Calcium Phosphate Nanocomposite Particles

In this study, the effect of silica/calcium phosphate (SiCaP) nanocomposite particles on the properties of a novel chitosan-based thermosensitive hydrogel system was examined. SiCaP nanocomposite powder was fabricated using a sol-gel method and then used to fabricate nanocomposite hydrogels (Ch- β/7.5SiCaP and Ch-β/15SiCaP) including chitosan and β-glycerophosphate (Ch-β) as a matrix. Results revealed that compared to the Ch-β hydrogel without SiCaP, the presence of SiCaP particles in nanocomposite hydrogels maintained pH stability during the sol-gel transition, accelerated the gelation and improved the stiffness of nanocomposite hydrogels. Gelation time at 37℃ was reduced approximately 75% and stiffness was increased approximately 115%. Both of these changes are attributed to chemical and physical interactions of the SiCaP bioactive particles with chitosan. Furthermore, compared to the Ch-β hydrogel, the presence of SiCaP in the Ch-β/7.5SiCaP nanocomposite hydrogel did not affect biocompatibility negatively, but improved osteoblastic cell differentiation. Our studies suggest that these nanocomposite hydrogels may offer an innovative approach to bone regeneration strategies.

Chitosan-Based Thermo-Sensitive Hydrogel Loading Oyster Peptides for Hemostasis Application.

Uncontrolled massive hemorrhage is one of the principal causes of death in trauma emergencies. By using catechol-modified chitosan (CS-C) as the matrix material and β glycerol phosphate (β-GP) as a thermo-sensitive agent, chitosan-based thermo-sensitive hydrogel loading oyster peptides (CS-C/OP/β-GP) were prepared at physiological temperature. The hemostatic performance of CS-C/OP/β-GP hydrogel was tested in vivo and in vitro, and its biological safety was evaluated. The results showed that the in vitro coagulation time and blood coagulation index of CS-C/OP/β-GP hydrogel were better than those of a commercial gelatin sponge. Notably, compared with the gelatin sponge, CS-C/OP/β-GP hydrogel showed that the platelet adhesion and erythrocyte adsorption rates were 38.98% and 95.87% higher, respectively. Additionally, the hemostasis time in mouse liver injury was shortened by 19.5%, and the mass of blood loss in the mouse tail amputation model was reduced by 18.9%. The safety evaluation results demonstrated that CS-C/OP/β-GP had no cytotoxicity to L929 cells, and the hemolysis rates were less than 5% within 1 mg/mL, suggesting good biocompatibility. In conclusion, our results indicate that CS-C/OP/β-GP is expected to be a promising dressing in the field of medical hemostasis.

A three dimensional in vivo model of breast cancer using a thermosensitive chitosan-based hydrogel and 4 T1 cell line in Balb/c.

The two-dimensional (2D) models of breast cancer still exhibit a limited success. Whereas, three-dimensional (3D) models provide more similar conditions to the tumor for growth of cancer cells. In this regard, a 3D in vivo model of breast cancer using 4 T1 cells and chitosan-based thermosensitive hydrogel were designed. Chitosan/β-glycerol phosphate hydrogel (Ch/β-GP) was prepared with a final ratio of 2% and 10%. The hydrogel properties were examined by Fourier transformed infrared spectroscopy, MTT assay, pH, scanning electron microscopy, and biodegradability assay. 3D model of breast cancer was induced by injection of 1 × 106 4 T1 cells in 100 μl hydrogel and 2D model by injection of 1 × 106 4 T1 cells in 100 μl phosphate-buffered saline (PBS) subcutaneously. After 3 weeks, induced tumors were evaluated by size and weight determination, ultrasound, hematoxylin- and eosin and Masson's trichrome staining and evaluating of cancer stem cells with CD44 and CD24 markers. The results showed that hydrogel with physiological pH had no cytotoxicity. In 3D model, tumor size and weight increased significantly (p ≤ .001) in comparison with 2D model. Histological and ultrasound analysis showed that 3D tumor model was more similar to breast cancer. Expression of CD44 and CD24 markers in the 3D model was more than 2D model (p ≤ .001). This 3D in vivo model of breast cancer mimicked native tumor and showed malignant tissue properties. Therefore, the use of such models can be effective in various cancer studies, especially in the field of cancer stem cells.