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1.
Clin Exp Dent Res ; 10(4): e932, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38973200

ABSTRACT

OBJECTIVES: There is a growing evidence to suggest augmenting peri-implant keratinized mucosa in the presence of ≤ 2 mm of keratinized mucosa. However, the most appropriate surgical technique and augmentation materials have yet to be defined. The aim of this systematic review and meta-analyses was to evaluate the clinical and patient-reported outcomes of augmenting keratinized mucosa around implants using free gingival graft (FGG) versus xenogeneic collagen matrix (XCM) before commencing prosthetic implant treatment. MATERIAL AND METHODS: Electronic databases were searched to identify observational studies comparing implant sites augmented with FGG to those augmented with XCM. The risk of bias was assessed using the Cochrane Collaboration's Risk of Bias tool. RESULTS: Six studies with 174 participants were included in the present review. Of these, 87 participants had FGG, whereas the remaining participants had XCM. At 6 months, sites augmented with FGG were associated with less changes in the gained width of peri-implant keratinized mucosa compared to those augmented with XCM (mean difference 1.06; 95% confidence interval -0.01 to 2.13; p = 0.05). The difference, however, was marginally significant. The difference between the two groups in changes in thickness of peri-implant keratinized mucosa at 6 months was statistically significantly in favor of FGG. On the other hand, XCM had significantly shorter surgical time, lower postoperative pain score, and higher color match compared to FGG. CONCLUSIONS: Within the limitation of this review, the augmentation of keratinized mucosa using FGG before the placement of the final prosthesis may have short-term positive effects on soft tissue thickness. XCM might be considered in aesthetically demanding implant sites and where patient comfort or shorter surgical time is a priority. The evidence support, however, is of low to moderate certainty; therefore, further studies are needed to support the findings of the present review.


Subject(s)
Collagen , Dental Implants , Gingiva , Humans , Collagen/therapeutic use , Gingiva/transplantation , Gingiva/pathology , Gingiva/surgery , Keratins , Mouth Mucosa/transplantation , Gingivoplasty/methods , Dental Implantation, Endosseous/methods , Heterografts
3.
Genes (Basel) ; 15(6)2024 May 24.
Article in English | MEDLINE | ID: mdl-38927617

ABSTRACT

Keratins are the main structural protein components of wool fibres, and variation in them and their genes (KRTs) is thought to influence wool structure and characteristics. The PCR-single strand conformation polymorphism technique has been used previously to investigate genetic variation in selected coding and intron regions of the type II sheep keratin gene KRT81, but no variation was identified. In this study, we used the same technique to explore the 5' untranslated region of KRT81 and detected three sequence variants (A, B and C) that contain four single nucleotide polymorphisms. Among the 389 Merino × Southdown cross sheep investigated, variant B was linked to a reduction in clean fleece weight, while C was associated with an increase in both greasy fleece weight and clean fleece weight. No discernible effects on staple length or mean-fibre-diameter-related traits were observed. These findings suggest that variation in ovine KRT81 might influence wool growth by changing the density of wool follicles in the skin, the density of individual fibres, or the area of the skin producing fibre, as opposed to changing the rate of extrusion of fibres or their diameter.


Subject(s)
Polymorphism, Single Nucleotide , Wool Fiber , Wool , Animals , Sheep/genetics , Sheep/growth & development , Wool/growth & development , Keratins, Type II/genetics , Keratins, Type II/metabolism , Keratins/genetics , Keratins/metabolism , Sheep, Domestic/genetics , Sheep, Domestic/growth & development
4.
Sci Rep ; 14(1): 12864, 2024 06 04.
Article in English | MEDLINE | ID: mdl-38834664

ABSTRACT

Natural polymer-based hydrogels have demonstrated great potential as wound-healing dressings. They help to maintain a moist wound environment as well as promote faster healing. In this work, a multifunctional hydrogel was prepared using keratin, sodium alginate, and carboxymethyl chitosan with tannic acid modification. Micro-morphology of hydrogels has been performed by scanning electron microscopy. Fourier Transform Infrared Spectroscopy reveals the presence of hydrogen bonding. The mechanical properties of the hydrogels were examined using a universal testing machine. Furthermore, we investigated several properties of the modified hydrogel. These properties include swelling rate, water retention, anti-freezing properties, antimicrobial and antioxidant properties, hemocompatibility evaluation and cell viability test in vitro. The modified hydrogel has a three-dimensional microporous structure, the swelling rate was 1541.7%, the elastic modulus was 589.74 kPa, the toughness was 211.74 kJ/m3, and the elongation at break was 75.39%, which was similar to the human skin modulus. The modified hydrogel also showed inhibition of S. aureus and E. coli, as well as a DPPH scavenging rate of 95%. In addition, the modified hydrogels have good biological characteristics. Based on these findings, the K/SA/CCS hydrogel holds promise for applications in biomedical engineering.


Subject(s)
Alginates , Chitosan , Hydrogels , Keratins , Tannins , Chitosan/chemistry , Chitosan/analogs & derivatives , Tannins/chemistry , Alginates/chemistry , Hydrogels/chemistry , Humans , Keratins/chemistry , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Staphylococcus aureus/drug effects , Antioxidants/chemistry , Antioxidants/pharmacology , Escherichia coli/drug effects , Wound Healing/drug effects , Cell Survival/drug effects , Spectroscopy, Fourier Transform Infrared , Elastic Modulus , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology
6.
Anim Sci J ; 95(1): e13969, 2024.
Article in English | MEDLINE | ID: mdl-38923230

ABSTRACT

We hypothesized that teats with a teat apex score (TAS) of 4 on a 4-point scale would exhibit elevated levels of denatured collagen compared with teats with lower TAS. We procured keratin layer and smooth muscle samples from Holsteins with TAS ranging from 1 to 4, as well as from crossbred heifers (Japanese Black male and Holstein female) with TAS of 1. Teats with a TAS of 4 demonstrated increased total collagen content, higher amounts of type I collagen (the harder, thicker variant), and reduced amounts of type III collagen (the softer, thinner variant) compared with teats with lower TAS. Teats with TAS of 3 and 4 exhibited evidence of damaged collagen in smooth muscle layers compared with teats with TAS of 1. Additionally, we identified 47-kDa heat shock protein-positive fibroblasts in the smooth muscles of teats with TAS of 3 and 4. Therefore, the smooth muscle of teats with a TAS of 4 exhibited increased amounts of denatured collagen in comparison to teats with lower TAS.


Subject(s)
Collagen , Keratins , Mammary Glands, Animal , Muscle, Smooth , Protein Denaturation , Animals , Cattle/metabolism , Female , Muscle, Smooth/metabolism , Collagen/metabolism , Collagen/analysis , Keratins/metabolism , Mammary Glands, Animal/metabolism , Male , Collagen Type I/metabolism , Collagen Type I/analysis , Fibroblasts/metabolism , Collagen Type III/metabolism , Collagen Type III/analysis
7.
Sci Rep ; 14(1): 13066, 2024 06 06.
Article in English | MEDLINE | ID: mdl-38844764

ABSTRACT

The aim of this study was to assess the surface and tissue quality of keratinized mucosa grafts (KMG) obtained using the conventional scalpel and mucotome techniques. This was an experimental in vitro/ex vivo study involving six porcine hemi-mandibles. Specimens were harvested using both the mucotome and conventional scalpel techniques, with randomization determining the choice of technique for tissue removal. The specimens were prepared following predefined laboratory protocols and subsequently subjected to optical microscopy for evaluating epithelial and connective tissue and scanning electron microscopy for topographical and 3D profilometry analysis. Tissues harvested using the mucotome exhibited a linear base and uniform thickness, along with the presence of submucosa and fibrous connective tissue, all of which are ideal for graft success. Differences in the surface characteristics of specimens obtained through the two techniques were observed during a comparative analysis of images obtained through both microscopy types. KMG obtained using the mucotome technique displayed greater uniformity and reduced undesirable cell presence compared to the scalpel technique, thereby enhancing the likelihood of success in soft tissue graft surgical procedures. This study provides valuable insights to oral healthcare professionals and may contribute to future research aimed at achieving more successful surgeries, shorter postoperative recovery times, reduced discomfort, and an overall more positive patient experience.


Subject(s)
Mandible , Mouth Mucosa , Animals , Swine , Mouth Mucosa/transplantation , Mouth Mucosa/cytology , Mandible/surgery , Keratins/metabolism , Microscopy, Electron, Scanning , Tissue and Organ Harvesting/methods
8.
J Microorg Control ; 29(2): 63-73, 2024.
Article in English | MEDLINE | ID: mdl-38880618

ABSTRACT

Cutibacterium acnes is an opportunistic pathogen recognized as a contributing factor to acne vulgaris. The accumulation of keratin and sebum plugs in hair follicles facilitates C. acnes proliferation, leading to inflammatory acne. Although numerous antimicrobial cosmetic products for acne-prone skin are available, their efficacy is commonly evaluated against planktonic cells of C. acnes. Limited research has assessed the antimicrobial effects on microorganisms within keratin and sebum plugs. This study investigates whether an antibacterial toner can penetrate keratin and sebum plugs, exhibiting bactericidal effects against C. acnes. Scanning electron microscopy and next-generation sequencing analysis of the keratin and sebum plug suggest that C. acnes proliferate within the plug, predominantly in a biofilm-like morphology. To clarify the potential bactericidal effect of the antibacterial toner against C. acnes inside keratin and sebum plugs, we immersed the plugs in the toner, stained them with LIVE/DEAD BacLight Bacterial Viability Kit to visualize microorganism viability, and observed them using confocal laser scanning microscopy. Results indicate that most microorganisms in the plugs were killed by the antibacterial toner. To quantitatively evaluate the bactericidal efficacy of the toner against C. acnes within keratin and sebum, we immersed an artificial plug with inoculated C. acnes type strain and an isolate collected from acne-prone skin into the toner and obtained viable cell counts. The number of the type strain and the isolate inside the artificial plug decreased by over 2.2 log and 1.2 log, respectively, showing that the antibacterial toner exhibits bactericidal effects against C. acnes via keratin and sebum plug penetration.


Subject(s)
Acne Vulgaris , Anti-Bacterial Agents , Keratins , Sebum , Sebum/metabolism , Anti-Bacterial Agents/pharmacology , Humans , Keratins/metabolism , Acne Vulgaris/microbiology , Acne Vulgaris/drug therapy , Biofilms/drug effects , Microbial Viability/drug effects , Propionibacteriaceae/drug effects , Propionibacteriaceae/metabolism , Propionibacteriaceae/genetics , Propionibacterium acnes/drug effects , Propionibacterium acnes/metabolism , Hair Follicle/microbiology , Hair Follicle/metabolism , Microscopy, Electron, Scanning
9.
Invest Ophthalmol Vis Sci ; 65(6): 28, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38888283

ABSTRACT

Purpose: The current study evaluated the lid margin microbiome of keratinized lid margins of patients with chronic Stevens-Johnson syndrome (SJS) and compared it with healthy controls and historically reported lid margin microbiome of patients with meibomian gland dysfunction (MGD). Methods: Eyelid margin swabs of 20 asymptomatic adults (mean age = 29 ± 12 years) and 10 patients with chronic SJS (mean age = 31.2 ± 14 years) with lid margin keratinization were sequenced using next generation of 16S rDNA V3 to V4 variable region. Within SJS, the keratinized lid margin microbiome was compared with adjacent eyelid skin. Results: All patients had obstructive MGD, and mean Schirmer I value was 2.8 ± 1.9 mm. The phyla were similar in two groups, whereas at the genera level, an increase in the relative abundance of Corynebacterium, Haemophilus, Azotobacter, and Afipia and a decrease of Acinetobacter was noted in SJS compared to healthy lid margins. SJS-associated microbiota displayed lesser diversity and more heterogeneity than healthy controls. The Principal Components Analysis (PCA) plot revealed wide separation in the SJS and the control groups. Correlational network analysis revealed Corynebacterium and Sphingomonas forming a major hub of negative interactions with other bacterial genera in the SJS group. Significant differences exist in the prevalent genera between keratinized lid margins and historically reported meibum microbiome of patients with MGD. In addition, the eyelid skin of patients with SJS had predominant Staphylococcus, whereas Corynebacterium and Pseudomonas were more in the keratinized lid margins compared to the eyelid skin microbiome. Conclusions: Lid margin microbiome is significantly altered in the keratinized lid margins of patients with SJS compared to the eyelid skin of patients with SJS, normal lid margins, and patients with MGD.


Subject(s)
Dry Eye Syndromes , Eyelids , Microbiota , Stevens-Johnson Syndrome , Humans , Male , Female , Adult , Dry Eye Syndromes/microbiology , Eyelids/microbiology , Stevens-Johnson Syndrome/microbiology , Middle Aged , Young Adult , Bacteria/genetics , Bacteria/isolation & purification , RNA, Ribosomal, 16S/genetics , DNA, Bacterial/genetics , DNA, Bacterial/analysis , Adolescent , Meibomian Glands/microbiology , Meibomian Glands/pathology , Meibomian Gland Dysfunction/microbiology , Keratins/metabolism
10.
Cell Death Dis ; 15(6): 439, 2024 Jun 21.
Article in English | MEDLINE | ID: mdl-38906852

ABSTRACT

Although adamantinomatous craniopharyngioma (ACP) is a tumour with low histological malignancy, there are very few therapeutic options other than surgery. ACP has high histological complexity, and the unique features of the immunological microenvironment within ACP remain elusive. Further elucidation of the tumour microenvironment is particularly important to expand our knowledge of potential therapeutic targets. Here, we performed integrative analysis of 58,081 nuclei through single-nucleus RNA sequencing and spatial transcriptomics on ACP specimens to characterize the features and intercellular network within the microenvironment. The ACP environment is highly immunosuppressive with low levels of T-cell infiltration/cytotoxicity. Moreover, tumour-associated macrophages (TAMs), which originate from distinct sources, highly infiltrate the microenvironment. Using spatial transcriptomic data, we observed one kind of non-microglial derived TAM that highly expressed GPNMB close to the terminally differentiated epithelial cell characterized by RHCG, and this colocalization was verified by asmFISH. We also found the positive correlation of infiltration between these two cell types in datasets with larger cohort. According to intercellular communication analysis, we report a regulatory network that could facilitate the keratinization of RHCG+ epithelial cells, eventually causing tumour progression. Our findings provide a comprehensive analysis of the ACP immune microenvironment and reveal a potential therapeutic strategy base on interfering with these two types of cells.


Subject(s)
Craniopharyngioma , Pituitary Neoplasms , Tumor Microenvironment , Humans , Craniopharyngioma/genetics , Craniopharyngioma/pathology , Craniopharyngioma/metabolism , Craniopharyngioma/immunology , Tumor Microenvironment/immunology , Pituitary Neoplasms/pathology , Pituitary Neoplasms/genetics , Pituitary Neoplasms/immunology , Pituitary Neoplasms/metabolism , Tumor-Associated Macrophages/metabolism , Tumor-Associated Macrophages/immunology , Male , Female , Keratins/metabolism , Transcriptome/genetics , Gene Expression Regulation, Neoplastic , Adult , Middle Aged , Multiomics
11.
Invest Ophthalmol Vis Sci ; 65(6): 37, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38935029

ABSTRACT

Purpose: To investigate the molecular mechanism of pathological keratinization in the chronic phase of ocular surface (OS) diseases. Methods: In this study, a comprehensive gene expression analysis was performed using oligonucleotide microarrays on OS epithelial cells obtained from three patients with pathological keratinization (Stevens-Johnson syndrome [n = 1 patient], ocular cicatricial pemphigoid [n = 1 patient], and anterior staphyloma [n = 1 patient]). The controls were three patients with conjunctivochalasis. The expression in some transcripts was confirmed using quantitative real-time PCR. Results: Compared to the controls, 3118 genes were significantly upregulated by a factor of 2 or more than one-half in the pathological keratinized epithelial cells (analysis of variance P < 0.05). Genes involved in keratinization, lipid metabolism, and oxidoreductase were upregulated, while genes involved in cellular response, as well as known transcription factors (TFs), were downregulated. Those genes were further analyzed with respect to TFs and retinoic acid (RA) through gene ontology analysis and known reports. The expression of TFs MYBL2, FOXM1, and SREBF2, was upregulated, and the TF ELF3 was significantly downregulated. The expression of AKR1B15, RDH12, and CRABP2 (i.e., genes related to RA, which is known to suppress keratinization) was increased more than twentyfold, whereas the expression of genes RARB and RARRES3 was decreased by 1/50. CRABP2, RARB, and RARRES3 expression changes were also confirmed by qRT-PCR. Conclusions: In pathological keratinized ocular surfaces, common transcript changes, including abnormalities in vitamin A metabolism, are involved in the mechanism of pathological keratinization.


Subject(s)
Gene Expression Regulation , Real-Time Polymerase Chain Reaction , Humans , Female , Male , Aged , Middle Aged , Oligonucleotide Array Sequence Analysis , Gene Expression Profiling , Pemphigoid, Benign Mucous Membrane/genetics , Pemphigoid, Benign Mucous Membrane/metabolism , Keratins/metabolism , Keratins/genetics , Corneal Diseases/genetics , Corneal Diseases/metabolism , Corneal Diseases/pathology , Epithelial Cells/metabolism , Epithelial Cells/pathology , Conjunctival Diseases/genetics , Conjunctival Diseases/metabolism , Conjunctival Diseases/pathology
12.
Anticancer Res ; 44(7): 3105-3113, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38925804

ABSTRACT

BACKGROUND/AIM: Classical serum cancer biomarkers, such as carcinoembryonic antigen (CEA) and cancer antigen 19-9 (CA 19-9), remain important tools in colorectal cancer (CRC) management for disease follow up. However, their sensitivity and specificity are low for diagnostic and prognostic evaluation. The aim of this study was to evaluate the potential of biomarkers reflecting biological activity of tumors - tissue polypeptide specific antigen (TPS), cytokeratin fragment 19 (CYFRA 21-1), thymidine kinase (TK), insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein 3 (IGF-BP3) - together with the CEA and CA 19-9 in CRC diagnosis and prognosis. PATIENTS AND METHODS: This is a retrospective study including 148 CRC patients and 68 age-matched healthy subjects. Serum biomarkers were measured in pre-operative serum samples using immunoanalytical methods. The end-point for the diagnostic evaluation was the area under the receiving operating characteristic curve (AUC ROC) of the biomarkers. The end-point for the prognostic evaluation was overall survival. RESULTS: Serum levels of CEA, CA 19-9, TPS, and TK were significantly increased in CRC early-stage patients compared with healthy controls. Each of the studied biomarkers had AUC between 0.6 and 0.7. Analysis of survival demonstrated that the patients with CEA, CA 19-9, cytokeratin, and TK above optimal cut offs had significantly shorter survival. A multivariate analysis performed on all the study biomarkers resulted in the selection of CYFRA 21-1 as the best performing biomarker with hazard ratio 10.413. CONCLUSION: The combination of cytokeratins and thymidine kinase with classical cancer biomarkers enables the prediction of tumor aggressiveness and long-term prognosis.


Subject(s)
Biomarkers, Tumor , CA-19-9 Antigen , Carcinoembryonic Antigen , Colorectal Neoplasms , Thymidine Kinase , Humans , Thymidine Kinase/blood , Colorectal Neoplasms/blood , Colorectal Neoplasms/diagnosis , Colorectal Neoplasms/pathology , Biomarkers, Tumor/blood , Male , Female , Aged , Middle Aged , Carcinoembryonic Antigen/blood , Retrospective Studies , Prognosis , CA-19-9 Antigen/blood , ROC Curve , Insulin-Like Growth Factor I/metabolism , Keratins/blood , Adult , Aged, 80 and over , Keratin-19/blood , Case-Control Studies , Antigens, Neoplasm/blood , Peptides
13.
Curr Microbiol ; 81(8): 227, 2024 Jun 16.
Article in English | MEDLINE | ID: mdl-38879855

ABSTRACT

Microbial degradation of keratin is characterized by its inherent safety, remarkable efficiency, and the production of copious degradation products. All these attributes contribute to the effective management of waste materials at high value-added and in a sustainable manner. Microbial degradation of keratin materials remains unclear, however, with variations observed in the degradation genes and pathways among different microorganisms. In this study, we sequenced the transcriptome of Purpureocillium lilacinum GZAC18-2JMP mycelia on control medium and the medium containing 1% feather powder, analyzed the differentially expressed genes, and revealed the degradation mechanism of chicken feathers by P. lilacinum GZAC18-2JMP. The results showed that the chicken feather degradation rate of P. lilacinum GZAC18-2JMP reached 64% after 216 h of incubation in the fermentation medium, reaching a peak value of 148.9 µg·mL-1 at 192 h, and the keratinase enzyme activity reached a peak value of 211 U·mL-1 at 168 h, which revealed that P. lilacinum GZAC18-2JMP had a better keratin degradation effect. A total of 1001 differentially expressed genes (DEGs) were identified from the transcriptome database, including 475 upregulated genes and 577 downregulated genes. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis of the DEGs revealed that the metabolic pathways related to keratin degradation were mainly sulfur metabolism, ABC transporters, and amino acid metabolism. Therefore, the results of this study provide an opportunity to gain further insight into keratin degradation and promote the biotransformation of feather wastes.


Subject(s)
Feathers , Hypocreales , Keratins , Transcriptome , Keratins/metabolism , Hypocreales/genetics , Hypocreales/metabolism , Animals , Feathers/metabolism , Chickens , Gene Expression Profiling , Fungal Proteins/genetics , Fungal Proteins/metabolism , Peptide Hydrolases/metabolism , Peptide Hydrolases/genetics , Mycelium/genetics , Mycelium/metabolism , Mycelium/growth & development , Fermentation , Biodegradation, Environmental
14.
Int J Mol Sci ; 25(11)2024 May 21.
Article in English | MEDLINE | ID: mdl-38891785

ABSTRACT

Intermediate filaments are one of three polymeric structures that form the cytoskeleton of epithelial cells. In the epithelium, these filaments are made up of a variety of keratin proteins. Intermediate filaments complete a wide range of functions in keratinocytes, including maintaining cell structure, cell growth, cell proliferation, cell migration, and more. Given that these functions are intimately associated with the carcinogenic process, and that hyperkeratinization is a quintessential feature of oral leukoplakias, the utility of keratins in oral leukoplakia is yet to be fully explored. This scoping review aims to outline the current knowledge founded on original studies on human tissues regarding the expression and utility of keratins as diagnostic, prognostic, and predictive biomarkers in oral leukoplakias. After using a search strategy developed for several scientific databases, namely, PubMed, Scopus, Web of Science, and OVID, 42 papers met the inclusion and exclusion criteria. One more article was added when it was identified through manually searching the list of references. The included papers were published between 1989 and 2024. Keratins 1-20 were investigated in the 43 included studies, and their expression was assessed in oral leukoplakia and dysplasia cases. Only five studies investigated the prognostic role of keratins in relation to malignant transformation. No studies evaluated keratins as a diagnostic adjunct or predictive tool. Evidence supports the idea that dysplasia disrupts the terminal differentiation pathway of primary keratins. Gain of keratin 17 expression and loss of keratin 13 were significantly observed in differentiated epithelial dysplasia. Also, the keratin 19 extension into suprabasal cells has been associated with the evolving features of dysplasia. The loss of keratin1/keratin 10 has been significantly associated with high-grade dysplasia. The prognostic value of cytokeratins has shown conflicting results, and further studies are required to ascertain their role in predicting the malignant transformation of oral leukoplakia.


Subject(s)
Keratins , Leukoplakia, Oral , Humans , Leukoplakia, Oral/metabolism , Leukoplakia, Oral/pathology , Leukoplakia, Oral/genetics , Keratins/metabolism , Keratins/genetics , Prognosis , Biomarkers, Tumor/metabolism
15.
J Colloid Interface Sci ; 669: 295-304, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38718583

ABSTRACT

Perovskite nanocrystals (PNCs) have emerged as promising candidates for fluorescent probes owing to their outstanding photoelectric properties. However, the conventional CsPbBr3 (CPB) NCs are extremely unstable in water, which has seriously limited their sensing applications in water environment. Herein, we present a powerful ligand engineering strategy for fabricating highly water-stable CPB NCs by using a biopolymer of wool keratin (WK) as the passivator and the polyaryl polymethylene isocyanate (PAPI) as the cross-linking agent. In particular, WK with multi-functional groups can serve as a polydentate ligand to firmly passivate CPB NCs by the ligand exchange process in hot toluene; and then the addition of PAPI can further encapsulate CPB NCs by the crosslinking reaction between PAPI and WK. Consequently, the as-prepared CPB/WK-PAPI NCs can maintain âˆ¼ 80 % of their relative photoluminescence (PL) intensity after 60 days in water, and they still maintain âˆ¼ 40 % of their relative PL intensity even after 512 days in the same environment, which is one of the best water stabilities compared previously reported polymer passivation methods. As a proof-of their application, the portable CPB/WK-PAPI NCs-based test strips are further developed as a fluorescent nanoprobe for real-time and visual monitoring amines and food freshness. Among various amine analytes, the as-prepared test strips exhibit higher sensitivity towards conjugated amines, achieving a remarkable detection limit of 18.3 nM for pyrrole. Our research not only introduces an innovative strategy involving natural biopolymers to enhance the water stability of PNCs, but also highlights the promising potential of PNCs for visually and portably detecting amines and assessing food freshness.


Subject(s)
Fluorescent Dyes , Keratins , Nanoparticles , Water , Wool , Nanoparticles/chemistry , Animals , Water/chemistry , Keratins/chemistry , Keratins/analysis , Wool/chemistry , Fluorescent Dyes/chemistry , Amines/chemistry , Particle Size , Surface Properties , Food Analysis/methods
16.
Cells ; 13(10)2024 May 09.
Article in English | MEDLINE | ID: mdl-38786031

ABSTRACT

The oral mucosa functions as a physico-chemical and immune barrier to external stimuli, and an adequate width of the keratinized mucosa around the teeth or implants is crucial to maintaining them in a healthy and stable condition. In this study, for the first time, bulk RNA-seq analysis was performed to explore the gene expression of laser microdissected epithelium and lamina propria from mice, aiming to investigate the differences between keratinized and non-keratinized oral mucosa. Based on the differentially expressed genes (DEGs) and Gene Ontology (GO) Enrichment Analysis, bone morphogenetic protein 2 (BMP-2) was identified to be a potential regulator of oral mucosal keratinization. Monoculture and epithelial-mesenchymal cell co-culture models in the air-liquid interface (ALI) indicated that BMP-2 has direct and positive effects on epithelial keratinization and proliferation. We further performed bulk RNA-seq of the ALI monoculture stimulated with BMP-2 in an attempt to identify the downstream factors promoting epithelial keratinization and proliferation. Analysis of the DEGs identified, among others, IGF2, ID1, LTBP1, LOX, SERPINE1, IL24, and MMP1 as key factors. In summary, these results revealed the involvement of a well-known growth factor responsible for bone development, BMP-2, in the mechanism of oral mucosal keratinization and proliferation, and pointed out the possible downstream genes involved in this mechanism.


Subject(s)
Bone Morphogenetic Protein 2 , Mouth Mucosa , Bone Morphogenetic Protein 2/metabolism , Bone Morphogenetic Protein 2/genetics , Mouth Mucosa/metabolism , Animals , Mice , Keratins/metabolism , Keratins/genetics , Cell Proliferation , Gene Expression Regulation , Humans , Gene Ontology
17.
Int J Biol Macromol ; 268(Pt 1): 131769, 2024 May.
Article in English | MEDLINE | ID: mdl-38692999

ABSTRACT

This study investigates the synthesis of selenium nanoparticles (SeNPs), owing to the low cost and abundance of selenium. However, the toxicity of SeNP prompts the development of a selenium nanocomposite (SeNC) containing pectin, keratin, and ferulic acid to improve the bioactivity of Se[0]. Further, incorporating the SeNC in a suitable formulation for drug delivery as a transdermal patch was worth studying. Accordingly, various analytical techniques were used to characterize the SeNPs and the SeNC, confirming successful synthesis and encapsulation. The SeNC exhibited notable particle size of 448.2 ± 50.2 nm, high encapsulation efficiency (98.90 % ± 2.4 %), 28.1 ± 0.45 drug loading, and sustained drug release at pH 5.5. Zeta potential and XPS confirmed the zero-oxidation state. The supramolecular structure was evident from spectral analysis endorsing the semi-crystalline nature of the SeNC and SEM images showcasing flower-shaped structures. Further, the SeNC demonstrated sustained drug release (approx. 22 % at 48 h) and wound-healing potential in L929 fibroblast cells. Subsequently, the SeNC loaded into a gelling agent exhibited shear thinning properties and improved drug release by nearly 58 %. A 3D printed reservoir-type transdermal patch was developed utilizing the SeNC-loaded gel, surpassing commercially available patches in characteristics such as % moisture uptake, tensile strength, and hydrophobicity. The patch, evaluated through permeation studies and CAM assay, exhibited controlled drug release and angiogenic properties for enhanced wound healing. The study concludes that this patch can serve as a smart dressing with tailored functionality for different wound stages, offering a promising novel drug delivery system for wound healing.


Subject(s)
Drug Liberation , Keratins , Nanogels , Pectins , Printing, Three-Dimensional , Selenium , Transdermal Patch , Selenium/chemistry , Pectins/chemistry , Keratins/chemistry , Animals , Nanogels/chemistry , Mice , Oxidation-Reduction , Wound Healing/drug effects , Cell Line , Nanocomposites/chemistry , Drug Carriers/chemistry , Drug Delivery Systems , Particle Size
18.
Sci Rep ; 14(1): 12374, 2024 05 29.
Article in English | MEDLINE | ID: mdl-38811642

ABSTRACT

Circulating tumor cells (CTCs) have gathered attention as a biomarker for carcinomas. However, CTCs in sarcomas have received little attention. In this work, we investigated cell surface proteins and antibody combinations for immunofluorescence detection of sarcoma CTCs. A microfluidic device that combines filtration and immunoaffinity using gangliosides 2 and cell surface vimentin (CSV) antibodies was employed to capture CTCs. For CTC detection, antibodies against cytokeratins 7 and 8 (CK), pan-cytokeratin (panCK), or a combination of panCK and CSV were used. Thirty-nine blood samples were collected from 21 patients of various sarcoma subtypes. In the independent samples study, samples were subjected to one of three antibody combination choices. Significant difference in CTC enumeration was found between CK and panCK + CSV, and between panCK and panCK + CSV. Upon stratification of CK+ samples, those of metastatic disease had a higher CTC number than those of localized disease. In the paired samples study involving cytokeratin-positive sarcoma subtypes, using panCK antibody detected more CTCs than CK. Similarly, for osteosarcoma, using panCK + CSV combination resulted in a higher CTC count than panCK. This study emphasized deliberate selection of cell surface proteins for sarcoma CTC detection and subtype stratification for studying cancers as heterogeneous as sarcomas.


Subject(s)
Biomarkers, Tumor , Neoplastic Cells, Circulating , Sarcoma , Humans , Neoplastic Cells, Circulating/pathology , Neoplastic Cells, Circulating/metabolism , Sarcoma/pathology , Sarcoma/blood , Sarcoma/diagnosis , Sarcoma/metabolism , Biomarkers, Tumor/blood , Female , Male , Membrane Proteins/metabolism , Membrane Proteins/immunology , Keratins/immunology , Keratins/metabolism , Middle Aged , Adult , Vimentin/metabolism , Vimentin/immunology , Aged , Antibodies/immunology , Cell Line, Tumor
19.
Int J Mol Sci ; 25(9)2024 May 03.
Article in English | MEDLINE | ID: mdl-38732209

ABSTRACT

One of the primary complications in generating physiologically representative skin tissue is the inability to integrate vasculature into the system, which has been shown to promote the proliferation of basal keratinocytes and consequent keratinocyte differentiation, and is necessary for mimicking representative barrier function in the skin and physiological transport properties. We created a 3D vascularized human skin equivalent (VHSE) with a dermal and epidermal layer, and compared keratinocyte differentiation (immunomarker staining), epidermal thickness (H&E staining), and barrier function (transepithelial electrical resistance (TEER) and dextran permeability) to a static, organotypic avascular HSE (AHSE). The VHSE had a significantly thicker epidermal layer and increased resistance, both an indication of increased barrier function, compared to the AHSE. The inclusion of keratin in our collagen hydrogel extracellular matrix (ECM) increased keratinocyte differentiation and barrier function, indicated by greater resistance and decreased permeability. Surprisingly, however, endothelial cells grown in a collagen/keratin extracellular environment showed increased cell growth and decreased vascular permeability, indicating a more confluent and tighter vessel compared to those grown in a pure collagen environment. The development of a novel VHSE, which incorporated physiological vasculature and a unique collagen/keratin ECM, improved barrier function, vessel development, and skin structure compared to a static AHSE model.


Subject(s)
Collagen , Hydrogels , Keratinocytes , Keratins , Skin , Humans , Hydrogels/chemistry , Collagen/chemistry , Collagen/metabolism , Keratinocytes/metabolism , Keratinocytes/cytology , Skin/metabolism , Skin/blood supply , Keratins/metabolism , Cell Differentiation , Cell Proliferation , Tissue Engineering/methods , Extracellular Matrix/metabolism , Cells, Cultured
20.
BMC Oral Health ; 24(1): 634, 2024 May 29.
Article in English | MEDLINE | ID: mdl-38811896

ABSTRACT

BACKGROUND: The aim of this study was to assess the outcomes of the combination technique of strip free gingival grafts (SFGG) and xenogeneic collagen matrix (XCM) in augmenting the width of keratinized mucosa (KMW) around dental implants, and compare its efficacy with the historical control group (FGG). METHODS: Thirteen patients with at least one site with KMW ≤ 2 mm after implant surgery were included and received SFGG in combination with XCM. Another thirteen patients with the same inclusion and exclusion criteria from the previous trial received FGG alone. The same outcomes as the previous trial were evaluated. KMW, thickness of keratinized mucosa (KMT), gingival index (GI) and probing depth (PD) were measured at baseline, 2 and 6 months. Postoperative pain, patient satisfaction and aesthetic outcomes were also assessed. RESULTS: At 6 months after surgery, the combination technique could attain 3.3 ± 1.6 mm of KMW. No significant change could be detected in GI or PD at 6 months compared to those at 2 months (p > 0.05). The postoperative pain and patient satisfaction in VAS were 2.6 ± 1.2 and 9.5 ± 1.2. The total score of aesthetic outcomes was 3.8 ± 1.2. In the historical FGG group, 4.6 ± 1.6 mm of KMW was reported at 6 months, and the total score of aesthetic outcomes was higher than the combination technique (4.8 ± 0.7 vs. 3.8 ± 1.2, p < 0.05). CONCLUSIONS: The combination technique of SFGG and XCM could increase KMW and maintain peri-implant health. However, this combination technique was associated with inferior augmentation and aesthetic outcomes compared with FGG alone. TRIAL REGISTRATION: This clinical trial was registered in the Chinese Clinical Trial Registry with registration number ChiCTR2200057670 on 15/03/2022.


Subject(s)
Collagen , Dental Implants , Gingiva , Humans , Female , Male , Collagen/therapeutic use , Middle Aged , Gingiva/transplantation , Adult , Patient Satisfaction , Periodontal Index , Gingivoplasty/methods , Keratins , Esthetics, Dental , Treatment Outcome , Pain, Postoperative/etiology , Mouth Mucosa/transplantation
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