tnf α concentrations  (Cusabio)

Journal: Advanced Science

Article Title: An Off‐the‐Shelf Artificial Blood Clot Hydrogel Neutralizing Multiple Proinflammatory Mediators for Pro‐Regenerative Periodontitis Treatment

doi: 10.1002/advs.202504106

Figure Lengend Snippet: CCS‐RSF@PRP hydrogels scavenged inflammatory mediators of ROS and LPS. a) The ROS scavenging mechanism of CCS‐RSF@PRP hydrogel. b) H 2 O 2 scavenging efficiency of CCS‐RSF@PRP hydrogel with CCS concentration of 10, 15, and 20 mg mL −1 , respectively (n = 3). c) UV‐vis spectra of DPPH after incubation with CCS‐RSF@PRP hydrogels. d) DPPH scavenging capabilities of CCS‐RSF@PRP hydrogels (n = 3). e) Representative images of DCFH‐labeled GECs after receiving different treatments. f) Quantitative analysis of mean fluorescence intensity of DCFH (n = 3). Histogram of flow cytometry (g) and corresponding mean fluorescence intensity (h) of GECs after receiving different treatments (n = 3). i) Quantification of LPS neutralization by activated or unactivated PRP as a function of LPS amount (n = 3). j) Representative CLSM images of GECs treated by LPS and LPS with CCS‐RSF@PRP hydrogel (red: F‐actin; blue: cell nuclear). k) Cell viability of GECs cultured with LPS and LPS with CCS‐RSF@PRP hydrogel evaluated by CCK‐8 assay (n = 8). l) Concentration of inflammatory factors TNF‐α in the cell supernatant of GECs (n = 3). m) Flow cytometry analysis of CD206 and CD86 expression (gated on F4/80 + cells). n) Percentage of M1 (F4/80 + CD86 + ) type macrophages (n = 3).

Article Snippet: The cell viability was quantified using CCK‐8 assay kit and TNF‐α cytokine concentration in the supernatant was tested by ELISA (CUSABIO, CSB‐E09315h) as above.

Techniques: Concentration Assay, Incubation, Labeling, Fluorescence, Flow Cytometry, Neutralization, Cell Culture, CCK-8 Assay, Expressing

Journal: Advanced Science

Article Title: An Off‐the‐Shelf Artificial Blood Clot Hydrogel Neutralizing Multiple Proinflammatory Mediators for Pro‐Regenerative Periodontitis Treatment

doi: 10.1002/advs.202504106

Figure Lengend Snippet: CCS‐RSF@PRP hydrogels neutralized proinflammatory cytokines and harnessed macrophages polarization toward M2 phenotype. a) Schematic diagram of macrophages polarization toward M2 phenotype induced by CCS‐RSF@PRP hydrogels. Neutralization of proinflammatory cytokines, including b) TNF‐α, c) IFN‐γ, and d) IL‐1β by CCS‐RSF@PRP hydrogel (n = 3). e) Representative CLSM images of BMDMs treated by IL‐4, CCS‐RSF, PRP, or CCS‐RSF@PRP hydrogel for 48 h, (red: CD206; green: F‐actin; blue: cell nuclear). f) The elongation of BMDMs (n = 3). g) Flow cytometry analysis of CD206 and MHC II expression (gated on F4/80 + cells). Percentage of h) M2 (F4/80 + CD206 + ) and i) M1 (F4/80 + MHC II + ) type macrophages (n = 3). Relative mRNA expression of j) CD206, k) Arg‐1, and l) VEGF‐A (n = 5). m, n) Protein expression level of Arg‐1 and iNOS in treated macrophages determined by western blot (n = 3).

Article Snippet: The cell viability was quantified using CCK‐8 assay kit and TNF‐α cytokine concentration in the supernatant was tested by ELISA (CUSABIO, CSB‐E09315h) as above.

Techniques: Neutralization, Flow Cytometry, Expressing, Western Blot

Journal: Advanced Science

Article Title: An Off‐the‐Shelf Artificial Blood Clot Hydrogel Neutralizing Multiple Proinflammatory Mediators for Pro‐Regenerative Periodontitis Treatment

doi: 10.1002/advs.202504106

Figure Lengend Snippet: CCS‐RSF@PRP hydrogels modulated the chronic inflammation microenvironment of periodontitis. a) Representative images of DHE staining (red: DHE, blue: DAPI), immunohistochemical staining of TNF‐α, immunofluorescence staining of CD206/CD68 and RUNX2 (red: RUNX2, blue: DAPI) on day 7 and day 28 after treatment. b) Statistical data of DHE positive area (%) (n = 3). c) Quantification of the relative content of TNF‐α in tissue (n = 3). d) Statistical data of the percentage of M2 macrophages (%) (n = 3). e) Statistical data of RUNX2 positive area (%) (n = 3). f‐h) Quantification of inflammatory cytokines LPS, IL‐1β, and TNF‐α in periodontium at the surgical site (n = 3). i‐j) Relative mRNA expression level of anti‐inflammatory cytokines (CD206, VEGF‐A, TGF‐β, and IL‐10) and pro‐inflammatory cytokines (TNF‐α and CD86).

Article Snippet: The cell viability was quantified using CCK‐8 assay kit and TNF‐α cytokine concentration in the supernatant was tested by ELISA (CUSABIO, CSB‐E09315h) as above.

Techniques: Staining, Immunohistochemical staining, Immunofluorescence, Expressing

Journal: Advanced Science

Article Title: An Off‐the‐Shelf Artificial Blood Clot Hydrogel Neutralizing Multiple Proinflammatory Mediators for Pro‐Regenerative Periodontitis Treatment

doi: 10.1002/advs.202504106

Figure Lengend Snippet: Mechanism analysis of periodontitis therapy with CCS‐RSF@PRP hydrogels. a) Venn diagram of the number of differentially expressed genes (DEGs). b) Volcano plots showing upregulated and downregulated DEGs. c) Gene ontology analysis of top 20 enriched terms between periodontitis group and CCS‐RSF@PRP hydrogels group. BP, biological processes; CC, cellular components; MF, molecular functions. d) KEGG pathway analysis demonstrating the top 13 signal pathways enriched by DEGs. e, f) Protein expression level of p‐MEK1, p‐Erk1/2, p‐IκBα, p‐NF‐κB and TNF‐α in periodontium of periodontitis and CCS‐RSF@PRP treatment group determined by western blot (n = 3). g, h) Protein expression level of p‐MEK1, p‐Erk1/2, p‐IκBα, p‐NF‐κB and TNF‐α of BMDMs in the control, TNF‐α, TNF‐α+infliximab+CCS‐RSF@PRP and TNF‐α+CCS‐RSF@PRP groups determined by western blot (n = 3). i) The mechanism of CCS‐RSF@PRP hydrogel remitting inflammation of periodontitis through TNF signaling pathway.

Article Snippet: The cell viability was quantified using CCK‐8 assay kit and TNF‐α cytokine concentration in the supernatant was tested by ELISA (CUSABIO, CSB‐E09315h) as above.

Techniques: Expressing, Western Blot, Control