In this work, we investigated the effect of the size and the chemical structure of crosslinkers on the properties of hyaluronic acid-based hydrogels prepared via an inverse electron demand ...Diels-Alder reaction. Hydrogels having loose and dense networks were designed by cross-linkers with and without polyethylene glycol (PEG) spacers of different molecular weights (1000 and 4000 g/mol). The study showed that the properties of hydrogels such as swelling ratios (20–55 times), morphology, stability, mechanical strength (storage modulus in the range 175–858 Pa), and drug loading efficiency (87 % ~ 90 %) were greatly influenced by the addition of PEG and changing its molecular weight in the cross-linker. Particularly, the presence of PEG chains in redox- responsive crosslinkers increased the doxorubicin release (85 %, after 168 h) and the degradation rate (96 %, after 10 d) of hydrogels in the simulated reducing medium (10 mM DTT). The in vitro cytotoxicity experiments conducted for HEK-293 cells revealed that the formulated hydrogels were biocompatible, which could be a promising candidate for drug delivery applications.
The objective of this study was to investigate inhibitory effects of a bioactive compound isolated from Ecklonia cava on fibrotic responses to transforming growth factor-β1 (TGF-β1)-stimulated Hs680. ...Tr human tracheal fibroblasts and the associated mechanisms of action. Post consecutive purification, a potent bioactive compound was identified phlorofucofuroeckol A. Phlorofucofuroeckol A significantly suppressed protein expression levels of collagen type I and α-smooth muscle actin (α-SMA) on TGF-β1-stimulated Hs680. Tr human tracheal fibroblasts. Further mechanistic studies determined that phlorofucofuroeckol A suppressed the phosphorylation of p38, extracellular regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) and SMAD 2/3 in TGF-β1-stimulated Hs680. Tr human tracheal fibroblasts. Moreover, we could show that phlorofucofuroeckol A inhibits binding of TGF-β1 to its TGF-β receptor by molecular docking. Based on the results, we propose that phlorofucofuroeckol A suppresses the MAPKs and SMAD 2/3 pathways and relieves cellular fibrotic activities, thus preventing tracheal fibrosis.
•This study was to purify phlorofucofuroeckol A from brown algae, Ecklonia cava.•Phlorofucofuroeckol A suppresses fibrotic markers in human tracheal fibroblasts.•Phlorofucofuroeckol A inhibits fibrosis via blocking TGF-β receptor.•Phlorofucofuroeckol A can be useful as an effective anti-fibrotic agent.
Transparent, biodegradable, mechanically robust, and surface‐patterned silk films were evaluated for the effect of surface morphology on human corneal fibroblast (hCF) cell proliferation, ...orientation, and ECM deposition and alignment. A series of dimensionally different surface groove patterns were prepared from optically graded glass substrates followed by casting poly(dimethylsiloxane) (PDMS) replica molds. The features on the patterned silk films showed an array of asymmetric triangles and displayed 37–342 nm depths and 445–3 582 nm widths. hCF DNA content on all patterned films were not significantly different from that on flat silk films after 4 d in culture. However, the depth and width of the grooves influenced cell alignment, while the depth differences affected cell orientation; overall, deeper and narrower grooves induced more hCF orientation. Over 14 d in culture, cell layers and actin filament organization demonstrated that confluent hCFs and their cytoskeletal filaments were oriented along the direction of the silk film patterned groove axis. Collagen type V and proteoglycans (decorin and biglycan), important markers of corneal stromal tissue, were highly expressed with alignment. Understanding corneal stromal fibroblast responses to surface features on a protein‐based biomaterial applicable in vivo for corneal repair potential suggests options to improve corneal tissue mimics. Further, the approaches provide fundamental biomaterial designs useful for bioengineering oriented tissue layers, an endemic feature in most biological tissue structures that lead to critical tissue functions.
We evaluated the effect of surface morphology on human corneal fibroblast cell proliferation, orientation and ECM deposition and alignment using surface‐patterned silk films. The depth and width of silk surface patterns affected cell alignment but not cell proliferation. Important ECM markers of corneal stromal tissue were highly expressed with alignment on the surface patterned silk films.
In this study, diselenide (Se-Se) and disulfide (S-S) redox-responsive core-cross-linked (CCL) micelles were synthesized using poly(ethylene oxide)
-
-poly(furfuryl methacrylate)
(PEO
-
-PFMA
), and ...their redox sensitivity was compared. A single electron transfer-living radical polymerization technique was used to prepare PEO
-
-PFMA
from FMA monomers and PEO
-Br initiators. An anti-cancer drug, doxorubicin (DOX), was incorporated into PFMA hydrophobic parts of the polymeric micelles, which were then cross-linked with maleimide cross-linkers, 1,6-bis(maleimide) hexane, dithiobis(maleimido) ethane and diselenobis(maleimido) ethane via Diels-Alder reaction. Under physiological conditions, the structural stability of both S-S and Se-Se CCL micelles was maintained; however, treatments with 10 mM GSH induced redox-responsive de-cross-linking of S-S and Se-Se bonds. In contrast, the S-S bond was intact in the presence of 100 mM H
O
while the Se-Se bond underwent de-crosslinking upon the treatment. DLS studies revealed that the size and PDI of (PEO
-
-PFMA
-Se)
micelles varied more significantly in response to changes in the redox environment than (PEO
-
-PFMA
-S)
micelles. In vitro release studies showed that the developed micelles had a lower drug release rate at pH 7.4, whereas a higher release was observed at pH 5.0 (tumor environment). The micelles were non-toxic against HEK-293 normal cells, which revealed that they could be safe for use. Nevertheless, DOX-loaded S-S/Se-Se CCL micelles exhibited potent cytotoxicity against BT-20 cancer cells. Based on these results, the (PEO
-
-PFMA
Se)
micelles can be more sensitive drug carriers than (PEO
-
-PFMA
-S)
micelles.
Display omitted
•NIR and reduction-responsive, gelatin-based hydrogels were developed by using norbornene-tetrazine click chemistry.•Water soluble diselenide-based crosslinkers were employed to ...prepare the bioorthogonal hydrogels.•Dual-responsive hydrogels exhibited quick followed by sustained release of DOX up to 48 h.
NIR and reduction-responsive gelatin hydrogels were designed for anti-tumor drug delivery application. Gelatin was functionalized with norbornene (Gel-Nb), followed by covalently cross-linking with a tetrazine (Tz)-based cross-linker (DSe-DPEG-DTz) possessing a redox-cleavable diselenide moiety. The resulting hydrogels were highly porous thanks to the N2 gas produced during the inverse electron demand Diels Alder ‘‘click reaction’’ between Nb and Tz. The hydrogels exhibited enhanced drug loading efficiency (≈ 94%) and excellent swelling ratios. The hydrogel prepared from the Nb:Tz mol. ratio of 10:10 (GHG-C) showed a storage modulus of 1100 Pa with an elastic rheological property. The doxorubicin (DOX)-loaded hydrogels (AR1) released minimal amounts (26%) of DOX at a physiological condition (PBS, pH 7.4). On the contrary, a fast release of DOX was observed in a reducing environment, where > 95% of DOX was released from AR3 after 48 h. The DOX and indocyanine green (ICG) co-loaded hydrogels (AR6) showed a burst release of DOX (>60% after 12 h) upon NIR irradiation, followed by a sustained release of the drug. The combined stimuli of GSH and NIR showed ≈ 85% in half of the total time. Gel-Nb, the cross-linker, and GHG-C were essentially non-toxic to the tested cell lines. Furthermore, AR3 and AR6 restricted the metabolic activities of BT-20 cells after treatment with GSH and NIR irradiation, respectively.
In anti‐cancer metastasis treatment, precise drug delivery to cancer cells remains a challenge. Innovative nanocomposites are developed to tackle these issues effectively. The approach involves the ...creation of manganese oxide (Mn3O4) nanoparticles (NPs) and their functionalization using trisodium citrate to yield functionalized Mn3O4 NPs (F‐Mn3O4 NPs), with enhanced water solubility, stability, and biocompatibility. Subsequently, the chemotherapeutic drug doxorubicin (DOX) is encapsulated with Mn3O4 NPs, resulting in DOX/Mn3O4 NPs. To achieve cell‐specific targeting, These NPs are coated with HeLa cell membranes (HCM), forming HCM/DOX/Mn3O4. For further refinement, a transferrin (Tf) receptor is integrated with cracked HCM to create Tf‐HCM/DOX/Mn3O4 nanocomposites (NC) with specific cell membrane targeting capabilities. The resulting Tf‐HCM/DOX/Mn3O4 NC exhibits excellent drug encapsulation efficiency (97.5%) and displays triggered drug release when exposed to NIR laser irradiation in the tumor's environment (pH 5.0 and 6.5). Furthermore, these nanocomposites show resistance to macrophage uptake and demonstrate homotypic cancer cell targeting specificity, even in the presence of other tumor cells. In vitro toxicity tests show that Tf‐HCM/DOX/Mn3O4 NC achieves significant anticancer activity against HeLa and BT20 cancer cells, with percentages of 76.46% and 71.36%, respectively. These results indicate the potential of Tf‐HCM/DOX/Mn3O4 NC as an effective nanoplatform for chemo‐photothermal therapy.
In anti‐cancer treatment, targeted nanocomposites are developed using Mn3O4 nanoparticles. Encapsulating doxorubicin within Mn3O4 NPs and coating them with HeLa cell membranes create Tf‐HCM/DOX/Mn3O4 nanocomposites, which release drugs when triggered by NIR laser. These nanocomposites resist macrophage uptake and specifically target homotypic cancer cells, even among other tumor cells. They hold significant potential for chemo‐photothermal therapy.
Stem cells are known to have excellent regenerative ability, which is primarily facilitated by indirect paracrine factors, rather than via direct cell replacement. The regenerative process is ...mediated by the release of extracellular matrix molecules, cytokines, and growth factors, which are also present in the media during cultivation. Herein, we aimed to demonstrate the functionality of key factors and mechanisms in skin regeneration through the analysis of conditioned media derived from fetal stem cells. A series of processes, including 3D pellet cultures, filtration and lyophilization is developed to fabricate human fetal cartilage-derived progenitor cells-conditioned media (hFCPCs-CM) and its useful properties are compared with those of human bone marrow-derived MSCs-conditioned media (hBMSCs-CM) in terms of biochemical characterization, and in vitro studies of fibroblast behavior, macrophage polarization, and burn wound healing. The hFCPCs-CM show to be devoid of cellular components but to contain large amounts of total protein, collagen, glycosaminoglycans, and growth factors, including IGFBP-2, IGFBP-6, HGF, VEGF, TGF β3, and M-CSF, and contain a specific protein, collagen alpha-1(XIV) compare with hBMSCs-CM. The therapeutic potential of hFCPCs-CM observes to be better than that of hBMSCs-CM in the viability, proliferation, and migration of fibroblasts, and M2 macrophage polarization in vitro, and efficient acceleration of wound healing and minimization of scar formation in third-degree burn wounds in a rat model. The current study shows the potential therapeutic effect of hFCPCs and provides a rationale for using the secretome released from fetal progenitor cells to promote the regeneration of skin tissues, both quantitatively and qualitatively.
Graphical abstract
The ready-to-use product of human fetal cartilage-derived progenitor cells-conditioned media (hFCPCs-CM) are fabricated via a series of techniques, including a 3D culture of hFCPCs, filtration using a 3.5 kDa cutoff dialysis membrane, and lyophilization of the CM. hFCPCs-CM contains many ECM molecules and biomolecules that improves wound healing through efficient acceleration of M2 macrophage polarization and reduction of scar formation.
This comprehensive review delves into the world of hyaluronic acid (HA) hydrogels, exploring their creation, characteristics, research methodologies, and uses. HA hydrogels stand out among natural ...polysaccharides due to their distinct features. Their exceptional biocompatibility makes them a top choice for diverse biomedical purposes, with a great ability to coexist harmoniously with living cells and tissues. Furthermore, their biodegradability permits their gradual breakdown by bodily enzymes, enabling the creation of temporary frameworks for tissue engineering endeavors. Additionally, since HA is a vital component of the extracellular matrix (ECM) in numerous tissues, HA hydrogels can replicate the ECM's structure and functions. This mimicry is pivotal in tissue engineering applications by providing an ideal setting for cellular growth and maturation. Various cross-linking techniques like chemical, physical, enzymatic, and hybrid methods impact the mechanical strength, swelling capacity, and degradation speed of the hydrogels. Assessment tools such as rheological analysis, electron microscopy, spectroscopy, swelling tests, and degradation studies are employed to examine their attributes. HA-based hydrogels feature prominently in tissue engineering, drug distribution, wound recovery, ophthalmology, and cartilage mending. Crafting HA hydrogels enables the production of biomaterials with sought-after qualities, offering avenues for advancements in the realm of biomedicine.
In this study, we have investigated whether human fetal cartilage progenitor cells (hFCPCs) have anti-inflammatory activity and can alleviate osteoarthritis (OA) phenotypes in vitro.
hFCPCs were ...stimulated with various cytokines and their combinations and expression of paracrine factors was examined to find an optimal priming factor. Human chondrocytes or SW982 synoviocytes were treated with interleukin-1β (IL-1β) to produce OA phenotype, and co-cultured with polyinosinic-polycytidylic acid (poly(I-C))-primed hFCPCs to address their anti-inflammatory effect by measuring the expression of OA-related genes. The effect of poly(I-C) on the surface marker expression and differentiation of hFCPCs into 3 mesodermal lineages was also examined.
Among the priming factors tested, poly(I-C) (1 µg/mL) most significantly induced the expression of paracrine factors such as indoleamine 2,3-dioxygenase, histocompatibility antigen, class I, G, tumor necrosis factor- stimulated gene-6, leukemia inhibitory factor, transforming growth factor-β1 and hepatocyte growth factor from hFCPCs. In the OA model in vitro, co-treatment of poly(I-C)-primed hFCPCs significantly alleviated IL-1β-induced expression of inflammatory factors such as IL-6, monocyte chemoattractant protein-1 and IL-1β, and matrix metalloproteinases in SW982, while it increased the expression of cartilage extracellular matrix such as aggrecan and collagen type II in human chondrocytes. We also found that treatment of poly(I-C) did not cause significant changes in the surface marker profile of hFCPCs, while showed some changes in the 3 lineages differentiation.
These results suggest that poly(I-C)-primed hFCPCs have an ability to modulate inflammatory response and OA phenotypes in vitro and encourage further studies to apply them in animal models of OA in the future.
The extracellular matrix (ECM) has many functions, such as segregating tissues, providing support, and regulating intercellular communication. Cartilage-derived ECM (CECM) can be prepared via ...consecutive processes of chemical decellularization and enzyme treatment. The purpose of this study was to improve and treat osteoarthritis (OA) using porcine knee articular CECM.
We assessed the rheological characteristics and pH of CECM solutions. Furthermore, we determined the effects of CECM on cell proliferation and cytotoxicity in the chondrocytes of New Zealand rabbits. The inhibitory effect of CECM on tumor necrosis factor (TNF)-α-induced cellular apoptosis was assessed using New Zealand rabbit chondrocytes and human synoviocytes. Finally, we examined the in vivo effects of CECM on inflammation control and cartilage degradation in an experimental OA-induced rat model. The rat model of OA was established by injecting monosodium iodoacetate into the intra-articular knee joint. The rats were then injected with CECM solution. Inflammation control and cartilage degradation were assessed by measuring the serum levels of proinflammatory cytokines and C-telopeptide of type II collagen and performing a histomorphological analysis.
CECM was found to be biocompatible and non-immunogenic, and could improve cell proliferation without inducing a toxic reaction. CECM significantly reduced cellular apoptosis due to TNF-α, significantly improved the survival of cells in inflammatory environments, and exerted anti-inflammatory effects.
Our findings suggest that CECM is an appropriate injectable material that mediates OA-induced inflammation.
PDF
