RESUMEN

Impedance aggregometry is an alternative to light transmission aggregometry that allows analysis of platelet function in whole blood samples. We hypothesized (1) impedance aggregometry would produce repeatable results, (2) inhibition of cyclooxygenase with aspirin would attenuate aggregation responses to collagen and abolish the aggregation response to arachidonic acid (AA), and (3) thromboxane receptor antagonism (terutroban) would attenuate the aggregation response to AA. Venous blood was obtained from 11 participants three times separated by at least 2 weeks. One sample followed 7-day-aspirin intervention (81 mg once daily; ASA), the others no intervention (control). Aggregation was induced using 1 µg/mL collagen ([col 1]), 5 µg/mL collagen ([col 5]), and 50 mM AA via impedance aggregometry to determine total aggregation (AUC) analyzed for intra-test repeatability, inter-test repeatability, intervention (ASA or control), and incubation (saline or terutroban). [col 1] showed high intra-test (p ≤ 0.03 visit 1 and 2) and inter-test repeatability (p < 0.01). [col 5] and AA showed intra- ([col 5] p < 0.01 visit 1 and 2; AA p < 0.001 visit 1 and 2) but not inter-test repeatability ([col 5] p = 0.48; AA p = 0.06). ASA attenuated AUC responses to [col 1] (p < 0.01), [col 5] (p = 0.03), and AA (p < 0.01). Terutroban attenuated AUC in response to AA (p < 0.01). [col 1] shows sufficient repeatability for longitudinal investigations of platelet function. [col 5] and AA may be used to investigate mechanisms of platelet function and metabolism at a single time point.

Asunto(s)

Aspirina , Inhibidores de la Ciclooxigenasa , Impedancia Eléctrica , Agregación Plaquetaria , Pruebas de Función Plaquetaria , Propionatos , Receptores de Tromboxanos , Humanos , Agregación Plaquetaria/efectos de los fármacos , Masculino , Proyectos Piloto , Femenino , Inhibidores de la Ciclooxigenasa/farmacología , Aspirina/farmacología , Receptores de Tromboxanos/antagonistas & inhibidores , Receptores de Tromboxanos/metabolismo , Adulto , Pruebas de Función Plaquetaria/métodos , Propionatos/farmacología , Naftalenos/farmacología , Ácido Araquidónico/farmacología , Plaquetas/efectos de los fármacos , Plaquetas/metabolismo , Persona de Mediana Edad , Inhibidores de Agregación Plaquetaria/farmacología , Colágeno/farmacología

RESUMEN

Additive manufacturing can develop regenerative scaffolds for wound healing. 3D printing offers meticulous porosity, mechanical integrity, cell adhesion and cost-effectiveness. Herein, we prepared ink composed of carboxymethyl cellulose (CMC), polyvinylpyrrolidone (PVP), collagen, and oregano extract for the fabrication of tissue constructs. The blend was optimized to form a homogeneous ink and rheological characterization demonstrated shear thinning behavior. The scaffolds were printed using Direct Ink Write (DIW) at a flow speed of 4 mm3/s and a layer height of 0.18 mm. The fabricated scaffolds demonstrated an ultimate tensile strength (UTS) and toughness of 730 KPa and 2.72 MJ/m3, respectively. Scanning Electron Microscopy (SEM) revealed an average pore size of 300 ± 30 µm. Fourier transform infrared spectroscopy (FTIR) analysis confirmed that all materials were present. The contact angle of the composite scaffold was 68° ± 1°. Moreover, the scaffolds presented 82 % mass loss (degradation) in phosphate buffer saline (PBS) over 14 days. The composite scaffold exhibited inhibition zones of 9 mm and 12 mm against Staphylococcus aureus and Escherichia coli, respectively. The PVP/CMC/collagen/oregano 3D printed scaffolds exhibited excellent biocompatibility with the mesenchymal stem cells and humman dermal fibroblast cells, confirmed by water-soluble tetrazolium - 8 (WST-8) assay (test conducted for 7 days). The enhanced angiogenic potential of said scaffold was assesed by release of vascular endothelial growth factor followed by further validation through in-vivo CAM assay. Thus, confirming suitability for the potential wound healing application.

Asunto(s)

Carboximetilcelulosa de Sodio , Colágeno , Origanum , Povidona , Andamios del Tejido , Cicatrización de Heridas , Povidona/química , Cicatrización de Heridas/efectos de los fármacos , Carboximetilcelulosa de Sodio/química , Carboximetilcelulosa de Sodio/farmacología , Andamios del Tejido/química , Colágeno/química , Colágeno/farmacología , Humanos , Origanum/química , Staphylococcus aureus/efectos de los fármacos , Impresión Tridimensional , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Escherichia coli/efectos de los fármacos , Tinta , Fibroblastos/efectos de los fármacos , Fibroblastos/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/citología , Antibacterianos/farmacología , Antibacterianos/química , Porosidad , Resistencia a la Tracción , Animales

RESUMEN

Liver sinusoidal endothelial cells (LSECs) are key targets for addressing metabolic dysfunction-associated steatotic liver disease (MASLD). However, isolating and culturing primary LSECs is challenging due to rapid dedifferentiation, resulting in loss of function. The extracellular matrix (ECM) likely plays a crucial role in maintaining the fate and function of LSECs. In this study, we explored the influence of liver-ECM (L-ECM) on liver cells and developed culture conditions that maintain the differentiated function of liver cells in vitro for prolonged periods. Porcine liver-derived L-ECM, containing 34.9 % protein, 0.045 % glycosaminoglycans, and negligible residual DNA (41.2 ng/mg), was utilized to culture primary rat liver cells in generated hydrogels. Proteomic analyses and molecular weight distribution of proteins of solubilized L-ECM revealed the typical diverse ECM core matrisome, with abundant collagens. L-ECM hydrogels showed suitable stiffness and stress relaxation properties. Furthermore, we demonstrated that collagen-rich L-ECM hydrogels enhanced LSECs' and hepatocytes' viability, and reduced the dedifferentiation rate of LSECs. In addition, hepatocyte function was maintained longer by culture on L-ECM hydrogels compared to traditional culturing. These beneficial effects are likely attributed to the bioactive macromolecules including collagens, and mechanical and microarchitectural properties of the L-ECM hydrogels.

Asunto(s)

Supervivencia Celular , Colágeno , Células Endoteliales , Matriz Extracelular , Hepatocitos , Hidrogeles , Hígado , Animales , Hidrogeles/química , Hidrogeles/farmacología , Hepatocitos/metabolismo , Hepatocitos/efectos de los fármacos , Hepatocitos/citología , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Ratas , Células Endoteliales/metabolismo , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Colágeno/química , Colágeno/farmacología , Colágeno/metabolismo , Hígado/metabolismo , Hígado/citología , Porcinos , Células Cultivadas , Masculino

RESUMEN

Corneal injuries play a significant role in global visual impairment, underscoring the demand for innovative biomaterials with specific attributes such as adhesion, cohesion, and regenerative potential. In this study, we have developed a biocompatible bioadhesive for corneal reconstruction. Derived from Collagen type I, naturally present in human corneal stromal tissue, the bioadhesive was cross-linked with modified polyethylene glycol diacrylate (PEGDA-DOPA), rendering it curable through visible light exposure and exhibiting superior adhesion to biological tissues even in wet conditions. The physicochemical characteristics of the proposed bioadhesive were customized by manipulating the concentration of its precursor polymers and adjusting the duration of photocrosslinking. To identify the optimal sample with maximum adhesion, mechanical strength, and biocompatibility, characterization tests were conducted. The optimal specimen, consisting of 30 % (w/v) PEGDA-DOPA and cured with visible light for 5 min, exhibited commendable adhesive strength of 783.6 kPa and shear strength of 53.7 kPa, surpassing that of commercialized eye adhesives.Additionally, biocompatibility test results indicated a notably high survival rate (>100 %) of keratocytes seeded on the hydrogel adhesive after 7 days of incubation. Consequently, this designed bioadhesive, characterized by high adhesion strength, robust mechanical strength, and excellent biocompatibility, is anticipated to enhance the spontaneous repair process of damaged corneal stromal tissue.

Asunto(s)

Perforación Corneal , Hidrogeles , Polietilenglicoles , Humanos , Hidrogeles/química , Hidrogeles/farmacología , Perforación Corneal/tratamiento farmacológico , Polietilenglicoles/química , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Colágeno/química , Colágeno/farmacología , Ensayo de Materiales , Adhesivos Tisulares/química , Adhesivos Tisulares/farmacología

RESUMEN

In this study, a UV-cured collagen-based film (C-P-H film) with high mechanical strength and antimicrobial properties was developed by riboflavin-mediated ultraviolet irradiation of collagen solution containing histidine-modified ε-polylysine. Fourier transform infrared analysis indicated that covalent cross-linking was formed between the collagen molecule and the histidine-grafted ε-polylysine. Compared with the pure collagen film, the C-P-H film containing 5 wt% histidine-modified ε-polylysine showed higher tensile strength (145.98 MPa), higher thermal denaturation temperature (76.5 °C), lower water vapor permeability (5.54 × 10-11 g m-1 s-1 Pa) and excellent antimicrobial activities against Escherichia coli and Staphylococcus aureus. In addition, the wrapping of the C-P-H film effectively inhibited bacterial growth of pork during storage time, successfully prolonging the shelf-life of pork by approximately 4 days compared to that of plastic wrap. These results suggested that collagen-based film grafted with histidine-modified ε-polylysine via riboflavin-mediated ultraviolet irradiation process had a great potential for pork preservation.

Asunto(s)

Colágeno , Escherichia coli , Embalaje de Alimentos , Conservación de Alimentos , Polilisina , Riboflavina , Staphylococcus aureus , Rayos Ultravioleta , Riboflavina/química , Riboflavina/farmacología , Animales , Colágeno/química , Colágeno/farmacología , Polilisina/química , Polilisina/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Porcinos , Embalaje de Alimentos/instrumentación , Conservación de Alimentos/instrumentación , Conservación de Alimentos/métodos , Resistencia a la Tracción , Antibacterianos/farmacología , Antibacterianos/química , Antiinfecciosos/farmacología , Antiinfecciosos/química

RESUMEN

The increasing cost of high-volume cultures and dependence on serum and growth factor supplementation limit the affordability of mesenchymal stromal cell (MSC) therapies. This has spurred interest in developing strategies that support adherent cell expansion while reducing raw material costs. Culture surfaces coated with sulfated glycosaminoglycans (GAGs), specifically heparan sulfate (HS), are an alternative to prolong growth factor retention in cell cultures. Unlike heparin, recombinant HS (rHS) offers strong binding affinity for multiple growth factors and extracellular matrix components, such as collagen I, without undesirable anticoagulant effects or xenobiotic health risks. The potential of rHS as a factor reservoir in MSC cultures remains underexplored. This study investigated the impact of rHS on the growth and anti-inflammatory properties of undifferentiated bone marrow MSCs in both planar and microcarrier-based cultures. It was hypothesized that rHS would enable MSC growth with minimal growth factor supplementation in a sulfation level-dependent manner. Cell culture surfaces were assembled via the layer-by-layer (LbL) method, combining alternating collagen I (COL) and rHS. These bilayers support cell adhesion and enable the incorporation of distinct sulfation levels on the culture surface. Examination of pro-mitogenic FGF and immunostimulatory IFN-γ release dynamics confirmed prolonged availability and sulfate level dependencies. Sulfated surfaces supported cell growth in low serum (2% FBS) and serum-free (SF) media at levels equivalent to standard culture conditions. Cell growth on rHS-coated surfaces in SF was comparable to that on heparin-coated surfaces and commercial surface-coated microcarriers in low serum. These growth benefits were observed in both planar and microcarrier (µCs) cultures. Additionally, rHS surfaces reduced ß-galactosidase expression relative to uncoated surfaces, delaying cell senescence. Multivariate analysis of cytokines in conditioned media indicated that rHS-containing surfaces enhanced cytokine levels relative to uncoated surfaces during IFN-γ stimulation and correlated with decreased pro-inflammatory macrophage activity. Overall, utilizing highly sulfated rHS with COL reduces the need for exogenous growth factors and effectively supports MSC growth and anti-inflammatory potency on planar and microcarrier surfaces under minimal factor supplementation.

Asunto(s)

Heparitina Sulfato , Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/efectos de los fármacos , Heparitina Sulfato/química , Heparitina Sulfato/farmacología , Heparitina Sulfato/metabolismo , Humanos , Técnicas de Cultivo de Célula/métodos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Adhesión Celular/efectos de los fármacos , Colágeno/química , Colágeno/farmacología , Colágeno/metabolismo , Medio de Cultivo Libre de Suero/química , Animales , Propiedades de Superficie

RESUMEN

Purpose: In this study, wound dressings were designed using zinc-modified marine collagen porous scaffold as host for wild bilberry (WB) leaves extract immobilized in functionalized mesoporous silica nanoparticles (MSN). These new composites were developed as an alternative to conventional wound dressings. In addition to the antibacterial activity of classic antibiotics, a polyphenolic extract could act as an antioxidant and/or an anti-inflammatory agent as well. Methods: Wild bilberry leaves extract was prepared by ultrasound-assisted extraction in ethanol and its properties were evaluated by UV-Vis spectroscopy (radical scavenging activity, total amount of polyphenols, flavonoids, anthocyanins, and condensed tannins). The extract components were identified by HPLC, and the antidiabetic properties of the extract were evaluated via α-glucosidase inhibitory activity. Spherical MSN were modified with propionic acid or proline moieties by post-synthesis method and used as carriers for the WB leaves extract. The textural and structural features of functionalized MSN were assessed by nitrogen adsorption/desorption isotherms, small-angle XRD, SEM, TEM, and FTIR spectroscopy. The composite porous scaffolds were prepared by freeze drying of the zinc-modified collagen suspension containing WB extract loaded silica nanoparticles. Results: The properties of the new composites demonstrated enhanced properties in terms of thermal stability of the zinc-collagen scaffold, without altering the protein conformation, and stimulation of NCTC fibroblasts mobility. The results of the scratch assay showed contributions of both zinc ions from collagen and the polyphenolic extract incorporated in functionalized silica in the wound healing process. The extract encapsulated in functionalized MSN proved enhanced biological activities compared to the extract alone: better inhibition of P. aeruginosa and S. aureus strains, higher biocompatibility on HaCaT keratinocytes, and anti-inflammatory potential demonstrated by reduced IL-1ß and TNF-α levels. Conclusion: The experimental data shows that the novel composites can be used for the development of effective wound dressings.

Asunto(s)

Vendajes , Colágeno , Nanopartículas , Extractos Vegetales , Hojas de la Planta , Dióxido de Silicio , Cicatrización de Heridas , Zinc , Extractos Vegetales/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Dióxido de Silicio/química , Dióxido de Silicio/farmacología , Colágeno/química , Colágeno/farmacología , Zinc/química , Zinc/farmacología , Nanopartículas/química , Cicatrización de Heridas/efectos de los fármacos , Antibacterianos/farmacología , Antibacterianos/química , Humanos , Andamios del Tejido/química , Animales , Antiinflamatorios/farmacología , Antiinflamatorios/química , Línea Celular , Porosidad , Fibroblastos/efectos de los fármacos , Antioxidantes/farmacología , Antioxidantes/química

RESUMEN

Periodontitis is a bacteria-induced chronic inflammatory disease characterized by degradation of the supporting tissue and bone in the oral cavity. Treatment modalities seek to facilitate periodontal rehabilitation while simultaneously preventing further gingival tissue recession and potentially bone atrophy. The aim of this study was to compare two differently sourced membranes, a resorbable piscine collagen membrane and a porcine-derived collagen membrane, in the repair of soft tissue defects utilizing a preclinical canine model. This in vivo component consisted of 10 beagles which were subjected to bilateral maxillary canine mucogingival flap defects, as well as bilateral soft tissue defects (or pouches) with no periodontal ligament damage in the mandibular canines. Defects received either a piscine-derived dermal membrane, (Kerecis® Oral, Ísafjörður, Iceland) or porcine-derived dermal membrane (Geistlich Mucograft®, Wolhusen, Switzerland) in a randomized fashion (to avoid site bias) and were allowed to heal for 30, 60, or 90 days. Statistical evaluation of tissue thickness was performed using general linear mixed model analysis of variance and least significant difference (LSD) post hoc analyses with fixed factors of time and membrane. Semi-quantitative analysis employed for inflammation assessment was evaluated using a chi-squared test along with a heteroscedastic t-test and values were reported as mean and corresponding 95% confidence intervals. In both the mucogingival flap defects and soft tissue gingival pouches, no appreciable qualitative differences were observed in tissue healing between the membranes. Furthermore, no statistical differences were observed in the thickness measurements between piscine- and porcine-derived membranes in the mucogingival flap defects (1.05 mm [±0.17] and 1.29 mm [±0.17], respectively [p = .06]) or soft tissue pouches (1.36 mm [±0.14] and 1.47 mm [±0.14], respectively [p = .27]), collapsed over time. Independent of membrane source (i.e., piscine or porcine), similar inflammatory responses were observed in both the maxilla and mandible at the three time points (p = .88 and p = .79, respectively). Histologic and histomorphometric evaluation results indicated that both membranes yielded equivalent tissue responses, remodeling dynamics and healing patterns for the mucogingival flap as well as the soft tissue gingival pouch defect models.

Asunto(s)

Colágeno , Cicatrización de Heridas , Animales , Perros , Porcinos , Colágeno/química , Colágeno/farmacología , Membranas Artificiales , Encía/patología

RESUMEN

Ovarian cancer, a malignant tumor that poses a significant threat to women's health, has seen a rise in incidence, prompting the urgent need for more effective treatment. This study primarily aimed to explore the potential of bovine collagen peptides in inhibiting ovarian cancer. The investigation in this study began with the identification of 268 peptide sequences through LC-MS/MS, followed by a screening process using molecular docking techniques to identify potential peptides capable of binding to EGFR. Subsequently, a series of experiments were performed, demonstrating the inhibitory effects of the peptide GPAGADGDRGEAGPAGPAGPAGPR on the proliferation of ovarian cancer cells. Transcriptomic analysis further revealed that this peptide can regulate cholesterol metabolism in ovarian cancer cells. Finally, a combination of time-resolved fluorescence resonance energy transfer, isothermal titration calorimetry, molecular docking, and molecular dynamics simulations were utilized to validate the ability of this peptide to bind to the epidermal growth factor receptor (EGFR) and impede the binding of epidermal growth factor (EGF) and EGFR.

Asunto(s)

Colágeno , Receptores ErbB , Simulación del Acoplamiento Molecular , Neoplasias Ováricas , Péptidos , Animales , Bovinos , Femenino , Humanos , Secuencia de Aminoácidos , Antineoplásicos/farmacología , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Colágeno/química , Colágeno/farmacología , Receptores ErbB/química , Simulación de Dinámica Molecular , Neoplasias Ováricas/tratamiento farmacológico , Péptidos/química , Péptidos/farmacología , Unión Proteica , Hidrolisados de Proteína/química , Hidrolisados de Proteína/farmacología

RESUMEN

Bone defects, resulting from trauma, inflammation, tumors, and various other factors, affect both health and quality of life. Although autologous bone transplantation is the gold-standard treatment for bone defects, it has disadvantages such as donor site limitations, prolonged surgical durations, and potential complications, necessitating the development of alternative bone tissue engineering materials. In this study, we used 3D printing technology to fabricate porous titanium implants characterized by superior biocompatibility and mechanical properties. Sodium alginate (SA) and strontium ions (Sr2+) were integrated into mineralized collagen matrices (MCs) to develop strontium-functionalized alginate-mineralized collagen hydrogels (SAMs) with high mechanical strength and sustained metal ion release ability. SAMs were seamlessly incorporated into the porous structures of 3D-printed titanium scaffolds, establishing a novel organic-inorganic bioactive interface. This composite system exhibited high biocompatibility in vitro and increased the expression of genes important for osteogenic differentiation and angiogenesis. In a rabbit model of femoral defect, the titanium implants effectively promoted bone and vascular regeneration on their surface, highlighting their potential in facilitating bone-implant integration.

Asunto(s)

Aleaciones , Colágeno , Hidrogeles , Oseointegración , Osteogénesis , Impresión Tridimensional , Andamios del Tejido , Titanio , Titanio/química , Animales , Osteogénesis/efectos de los fármacos , Hidrogeles/química , Hidrogeles/farmacología , Andamios del Tejido/química , Conejos , Oseointegración/efectos de los fármacos , Colágeno/química , Colágeno/farmacología , Porosidad , Aleaciones/química , Aleaciones/farmacología , Neovascularización Fisiológica/efectos de los fármacos , Estroncio/química , Estroncio/farmacología , Ingeniería de Tejidos/métodos , Alginatos/química , Alginatos/farmacología , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología

RESUMEN

Severe fracture non-union often accompanied by damaged or even absent periosteum remains a significant challenge. This paper presents a novel tri-layer bionic periosteum with gradient structure and mineralized collagen (MC) mimics natural periosteum for in-situ repair and bone regeneration. The construct with ultrasonic polylactic acid as the loose outer fibrous layer (UPLA), poly(ε-caprolactone) as the intermediate barrier layer (PCL-M), and poly(ε-caprolactone)/MC as the inner osteoblastic layer (PM) was prepared. The physicochemical properties of layers were investigated. UPLA/PCL-M/PM exhibited a tensile strength (3.55 ± 0.23 MPa) close to that of natural periosteum and excellent adhesion between the layers. In vitro experiments demonstrated that all layers had no toxicity to cells. UPLA promoted inward growth of mouse fibroblasts. PCL-M with a uniform pore size (2.82 ± 0.05 µm) could achieve a barrier effect against fibroblasts according to the live/dead assay. Meanwhile, PM could effectively promote cell migration with high alkaline phosphatase expression and significant mineralization of the extracellular matrix. Besides, in vivo experiments showed that UPLA/PCL-M/PM significantly promoted the regeneration of bone and early angiogenesis. Therefore, this construct with gradient structure developed in this paper would have great application potential in the efficient and high-quality treatment of severe fractures with periosteal defects.

Asunto(s)

Regeneración Ósea , Colágeno , Periostio , Poliésteres , Andamios del Tejido , Animales , Regeneración Ósea/efectos de los fármacos , Colágeno/química , Colágeno/farmacología , Ratones , Poliésteres/química , Andamios del Tejido/química , Regeneración Tisular Dirigida/métodos , Ingeniería de Tejidos/métodos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Fibroblastos/efectos de los fármacos

RESUMEN

In our search for a biocompatible composite hemostatic dressing, we focused on the design of a novel biomaterial composed of two natural biological components, collagen and sodium alginate (SA), cross-linked using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) and oxidized sodium alginate (OSA). We conducted a series of tests to evaluate the physicochemical properties, acute systemic toxicity, skin irritation, intradermal reaction, sensitization, cytotoxicity, and in vivo femoral artery hemorrhage model. The results demonstrated the excellent biocompatibility of the collagen/sodium alginate (C/SA)-based dressings before and after crosslinking. Specifically, the femoral artery hemorrhage model revealed a significantly shortened hemostasis time of 132.5 ± 12.82 s for the EDC/NHS cross-linked dressings compared to the gauze in the blank group (hemostasis time of 251.43 ± 10.69 s). These findings indicated that C/SA-based dressings exhibited both good biocompatibility and a significant hemostatic effect, making them suitable for biomedical applications.

Asunto(s)

Alginatos , Vendajes , Colágeno , Hemostáticos , Alginatos/química , Alginatos/farmacología , Animales , Colágeno/química , Colágeno/farmacología , Hemostáticos/química , Hemostáticos/farmacología , Ratones , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Ensayo de Materiales , Hemorragia/tratamiento farmacológico , Masculino , Ratas , Hemostasis/efectos de los fármacos , Arteria Femoral

RESUMEN

Collagen-based (COL) hydrogels could be a promising treatment option for injuries to the articular cartilage (AC) becuase of their similarity to AC native extra extracellular matrix. However, the high hydration of COL hydrogels poses challenges for AC's mechanical properties. To address this, we developed a hydrogel platform that incorporating cellulose nanocrystals (CNCs) within COL and followed by plastic compression (PC) procedure to expel the excessive fluid out. This approach significantly improved the mechanical properties of the hydrogels and enhanced the chondrogenic differentiation of mesenchymal stem cells (MSCs). Radially confined PC resulted in higher collagen fibrillar densities together with reducing fibril-fibril distances. Compressed hydrogels containing CNCs exhibited the highest compressive modulus and toughness. MSCs encapsulated in these hydrogels were initially affected by PC, but their viability improved after 7 days. Furthermore, the morphology of the cells and their secretion of glycosaminoglycans (GAGs) were positively influenced by the compressed COL-CNC hydrogel. Our findings shed light on the combined effects of PC and CNCs in improving the physical and mechanical properties of COL and their role in promoting chondrogenesis.

Asunto(s)

Diferenciación Celular , Celulosa , Condrogénesis , Colágeno , Hidrogeles , Células Madre Mesenquimatosas , Nanopartículas , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Celulosa/química , Celulosa/farmacología , Condrogénesis/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Nanopartículas/química , Colágeno/química , Colágeno/farmacología , Hidrogeles/química , Hidrogeles/farmacología , Animales , Plásticos/química , Plásticos/farmacología , Supervivencia Celular/efectos de los fármacos , Glicosaminoglicanos/metabolismo , Cartílago/citología , Cartílago/efectos de los fármacos

RESUMEN

BACKGROUND: Our objective was to conduct a systematic review of the effects of hydrolyzed collagen supplementation on the proliferation and activation of fibroblasts. METHODS: The search was conducted for journals that published articles in the English language, peer-reviewed, meeting the following criteria: (a) randomized clinical trials, (b) randomized studies in animals or humans, (c) in vitro studies, (d) studies using hydrolyzed collagens or collagen peptides, and (e) studies assessing alterations on fibroblasts as the primary or secondary outcome. We utilized the main journal databases PubMed/Web of Science and ongoing reviews by PROSPERO. For bias risk and methodological quality, we used an adaptation of the Downs and Black checklist. Our review followed the PRISMA checklist, conducted from February 2024 to the first week of March 2024, by two independent researchers (P.A.Q.I. and R.P.V.). RESULTS: Eleven studies were included in this review, where our findings reinforce the notion that hydrolyzed collagens or collagen peptides at concentrations of 50-500 µg/mL are sufficient to stimulate fibroblasts in human and animal tissues without inducing toxicity. Different enzymatic processes may confer distinct biological properties to collagens, allowing for scenarios favoring fibroblast promotion or antioxidant effects. Lastly, collagens with lower molecular weights exhibit greater bioavailability to adjacent tissues. CONCLUSIONS: Hydrolyzed collagens or collagen peptides with molecular sizes ranging from <3 to 3000 KDa promote the stimulation of fibroblasts in human tissues.

Asunto(s)

Colágeno , Suplementos Dietéticos , Fibroblastos , Colágeno/farmacología , Humanos , Fibroblastos/efectos de los fármacos , Animales , Proliferación Celular/efectos de los fármacos , Hidrólisis

RESUMEN

Developing a reliable method to predict thrombocytopenia is imperative in drug discovery. Here, we establish an assay using a microphysiological system (MPS) to recapitulate the in-vivo mechanisms of platelet aggregation and adhesion. This assay highlights the role of shear stress on platelet aggregation and their interactions with vascular endothelial cells. Platelet aggregation induced by soluble collagen was detected under agitated, but not static, conditions using a plate shaker and gravity-driven flow using MPS. Notably, aggregates adhered on vascular endothelial cells under gravity-driven flow in the MPS, and this incident increased in a concentration-dependent manner. Upon comparing the soluble collagen-induced aggregation activity in platelet-rich plasma (PRP) and whole blood, remarkable platelet aggregate formation was observed at concentrations of 30 µg/mL and 3 µg/mL in PRP and whole blood, respectively. Moreover, ODN2395, an oligonucleotide, induced platelet aggregation and adhesion to vascular endothelial cells. SYK inhibition, which mediated thrombogenic activity via glycoprotein VI on platelets, ameliorated platelet aggregation in the system, demonstrating that the mechanism of platelet aggregation was induced by soluble collagen and oligonucleotide. Our evaluation system partially recapitulated the aggregation mechanisms in blood vessels and can contribute to the discovery of safe drugs to mitigate the risk of thrombocytopenia.

Asunto(s)

Plaquetas , Agregación Plaquetaria , Trombocitopenia , Agregación Plaquetaria/efectos de los fármacos , Humanos , Trombocitopenia/inducido químicamente , Plaquetas/efectos de los fármacos , Plaquetas/metabolismo , Colágeno/metabolismo , Colágeno/farmacología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Adhesividad Plaquetaria/efectos de los fármacos , Quinasa Syk/metabolismo , Quinasa Syk/antagonistas & inhibidores , Plasma Rico en Plaquetas/metabolismo , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Sistemas Microfisiológicos

RESUMEN

Collagen's unique properties promise hemostatic potential, but its sponge form's stability and mechanics need improvement. In this study, we developed a series of homeostatic sponges by co-assembling collagen and curdlan at different ratios into hydrogels, followed by freeze-drying treatment. The incorporation of curdlan into collagen sponges has been found to significantly enhance the sponge's properties, including increased porosity, elevated water uptake, improved elasticity, and enhanced resistance to degradation. In vitro cytotoxicity and hemolysis assays have demonstrated the biocompatibility and nontoxicity of composite sponges. In mouse liver perforation and incision models, the composite sponges achieved rapid coagulation within 67 s and 75 s, respectively, outperforming gauze and gelatin sponge in reducing blood loss. Furthermore, composite sponges demonstrated superior wound healing potential in mice full-thickness skin defects model, with accelerated healing rates observed at days 3, 7, and 14 compared to the control group. Overall, collagen/curdlan composite sponge show promise for hemostasis and wound healing applications.

Asunto(s)

Colágeno , Hemostasis , Cicatrización de Heridas , beta-Glucanos , Animales , Cicatrización de Heridas/efectos de los fármacos , Colágeno/química , Colágeno/farmacología , beta-Glucanos/farmacología , beta-Glucanos/química , Ratones , Hemostasis/efectos de los fármacos , Piel/efectos de los fármacos , Piel/lesiones , Hidrogeles/química , Hidrogeles/farmacología , Hemólisis/efectos de los fármacos , Hemostáticos/farmacología , Hemostáticos/química , Porosidad , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Humanos , Masculino

RESUMEN

The potential of recombinant materials in the field of adipose tissue engineering (ATE) is investigated using a bottom-up tissue engineering (TE) approach. This study explores the synthesis of different photo-crosslinkable gelatin derivatives, including both natural and recombinant materials, with a particular emphasis on chain growth and step growth polymerization. Gelatin type B (Gel-B) and a recombinant collagen peptide (RCPhC1) are used as starting materials. The gel fraction and mass swelling properties of 2D hydrogel films are evaluated, revealing high gel fractions exceeding 94% and high mass swelling ratios >15. In vitro experiments with encapsulated adipose-derived stem cells (ASCs) indicate viable cells (>85%) throughout the experiment with the RCPhC1-based hydrogels showing a higher number of stretched ASCs. Triglyceride assays show the enhanced differentiation potential of RCPhC1 materials. Moreover, the secretome analysis reveal the production of adipose tissue-specific proteins including adiponectin, adipsin, lipocalin-2/NGAL, and PAL-1. RCPhC1-based materials exhibit higher levels of adiponectin and adipsin production, indicating successful differentiation into the adipogenic lineage. Overall, this study highlights the potential of recombinant materials for ATE applications, providing insights into their physico-chemical properties, mechanical strength, and cellular interactions.

Asunto(s)

Tejido Adiposo , Materiales Biocompatibles , Hidrogeles , Proteínas Recombinantes , Ingeniería de Tejidos , Tejido Adiposo/citología , Humanos , Ingeniería de Tejidos/métodos , Materiales Biocompatibles/farmacología , Materiales Biocompatibles/química , Hidrogeles/química , Hidrogeles/farmacología , Proteínas Recombinantes/farmacología , Gelatina/química , Gelatina/farmacología , Regeneración/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Células Madre/citología , Células Madre/efectos de los fármacos , Células Madre/metabolismo , Colágeno/química , Colágeno/farmacología , Adipogénesis/efectos de los fármacos

RESUMEN

Purpose: Surgical site infections pose a significant challenge for medical services. Systemic antibiotics may be insufficient in preventing bacterial biofilm development. With the local administration of antibiotics, it is easier to minimize possible complications, achieve drugs' higher concentration at the injured site, as well as provide their more sustained release. Therefore, the main objective of the proposed herein studies was the fabrication and characterization of innovative hydrogel-based composites for local vancomycin (VAN) therapy. Methods: Presented systems are composed of ionically gelled chitosan particles loaded with vancomycin, embedded into biomimetic collagen/chitosan/hyaluronic acid-based hydrogels crosslinked with genipin and freeze-dried to serve in a flake/disc-like form. VAN-loaded carriers were characterized for their size, stability, and encapsulation efficiency (EE) using dynamic light scattering technique, zeta potential measurements, and UV-Vis spectroscopy, respectively. The synthesized composites were tested in terms of their physicochemical and biological features. Results: Spherical structures with sizes of about 200 nm and encapsulation efficiencies reaching values of approximately 60% were obtained. It was found that the resulting particles exhibit stability over time. The antibacterial activity of the developed materials against Staphylococcus aureus was established. Moreover, in vitro cell culture study revealed that the surfaces of all prepared systems are biocompatible as they supported the proliferation and adhesion of the model MG-63 cells. In addition, we have demonstrated significantly prolonged VAN release while minimizing the initial burst effect for the composites compared to bare nanoparticles and verified their desired physicochemical features during swellability, and degradation experiments. Conclusion: It is expected that the developed herein system will enable direct delivery of the antibiotic at an exposed to infections surgical site, providing drugs sustained release and thus will reduce the risk of systemic toxicity. This strategy would both inhibit biofilm formation and accelerate the healing process.

Asunto(s)

Antibacterianos , Quitosano , Hidrogeles , Staphylococcus aureus , Vancomicina , Vancomicina/química , Vancomicina/farmacología , Vancomicina/administración & dosificación , Vancomicina/farmacocinética , Antibacterianos/química , Antibacterianos/farmacología , Antibacterianos/administración & dosificación , Hidrogeles/química , Hidrogeles/farmacología , Staphylococcus aureus/efectos de los fármacos , Humanos , Quitosano/química , Ácido Hialurónico/química , Ácido Hialurónico/farmacología , Portadores de Fármacos/química , Colágeno/química , Colágeno/farmacología , Tamaño de la Partícula , Liberación de Fármacos , Infección de la Herida Quirúrgica/prevención & control , Infección de la Herida Quirúrgica/tratamiento farmacológico , Pruebas de Sensibilidad Microbiana , Biopelículas/efectos de los fármacos

RESUMEN

Guided bone regeneration (GBR) membranes that reside at the interface between the bone and soft tissues for bone repair attract increasing attention, but currently developed GBR membranes suffer from relatively poor osteogenic and antibacterial effects as well as limited mechanical property and biodegradability. We present here the design and fabrication of a bifunctional Janus GBR membrane based on a shear flow-driven layer by a layer self-assembly approach. The Janus GBR membrane comprises a calcium phosphate-collagen/polyethylene glycol (CaP@COL/PEG) layer and a chitosan/poly(acrylic acid) (CHI/PAA) layer on different sides of a collagen membrane to form a sandwich structure. The membrane exhibits good mechanical stability and tailored biodegradability. It is found that the CaP@COL/PEG layer and CHI/PAA layer contribute to the osteogenic differentiation and antibacterial function, respectively. In comparison with the control group, the Janus GBR membrane displays a 2.52-time and 1.84-time enhancement in respective volume and density of newly generated bone. The greatly improved bone repair ability of the Janus GBR membrane is further confirmed through histological analysis, and it has great potential for practical applications in bone tissue engineering.

Asunto(s)

Antibacterianos , Regeneración Ósea , Osteogénesis , Regeneración Ósea/efectos de los fármacos , Antibacterianos/farmacología , Antibacterianos/química , Osteogénesis/efectos de los fármacos , Animales , Quitosano/química , Quitosano/farmacología , Fosfatos de Calcio/química , Fosfatos de Calcio/farmacología , Membranas Artificiales , Colágeno/química , Colágeno/farmacología , Polietilenglicoles/química , Polietilenglicoles/farmacología , Regeneración Tisular Dirigida/métodos , Ingeniería de Tejidos/métodos , Diferenciación Celular/efectos de los fármacos

RESUMEN

In this study, a series of collagen-chitosan-eugenol (CO-CS-Eu) flow-casting composite films were prepared using collagen from sturgeon skin, chitosan, and eugenol. The physicochemical properties, mechanical properties, microstructure, as well as antioxidant and antimicrobial activities of the composite membranes were investigated by various characterization techniques. The findings revealed that the inclusion of eugenol augmented the thickness of the film, darkened its color, reduced the transparency, and enhanced the ultraviolet light-blocking capabilities, with the physicochemical properties of the CO-CS-0.25%Eu film being notably favorable. Eugenol generates increasingly intricate matrices that disperse within the system, thereby modifying the optical properties of the material. Furthermore, the tensile strength of the film decreased from 70.97 to 20.32 MPa, indicating that eugenol enhances the fluidity and ductility of the film. Added eugenol also exhibited structural impact by loosening the film cross-section and decreasing its density. The Fourier transform infrared spectroscopy results revealed the occurrence of several intermolecular interactions among collagen, chitosan, and eugenol. Moreover, the incorporation of eugenol bolstered the antioxidant and antimicrobial capabilities of the composite film. This is primarily attributed to the abundant phenolic/hydroxyl groups present in eugenol, which can react with free radicals by forming phenoxy groups and neutralizing hydroxyl groups. Consequently, inclusion of eugenol substantially enhances the freshness retention performance of the composite film. PRACTICAL APPLICATION: ● The CO-CS-Eu film utilizes collagen from sturgeon skin, improving the use of sturgeon resources.● Different concentrations of eugenol altered its synergistic effect with chitosan.● The CO-CS-Eu film is composed of natural products with safe and edible properties.

Asunto(s)

Antioxidantes , Quitosano , Colágeno , Eugenol , Peces , Piel , Resistencia a la Tracción , Eugenol/farmacología , Eugenol/química , Quitosano/química , Quitosano/farmacología , Animales , Colágeno/química , Colágeno/farmacología , Piel/efectos de los fármacos , Piel/química , Antioxidantes/farmacología , Antioxidantes/química , Embalaje de Alimentos/métodos , Antiinfecciosos/farmacología , Antiinfecciosos/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos