Journal: Scientific Reports

Article Title: Various distribution of BMPs in different periosteal layers contributing to inconsistent osteoinductivity of DBM-based products

doi: 10.1038/s41598-026-35269-z

Figure Lengend Snippet: The amount of BMP-2 in DBM significantly increased in the order of OD > MD > ID using either extracted method, and was more than cortical particles.

Article Snippet: Sandwich enzyme-linked immunosorbent assay (ELISA) was used to assess the amounts of BMP with Commercially available quantikine kits purchased from MULTI SCIENCES (LIANKE) BIOTECH, CO, LTD (human BMP-2 ELISA Kit, sensitivity, 1.73 pg/ml; BMP-7 ELISA Kit, sensitivity, 2.06 pg/ml).

Techniques:

Journal: Scientific Reports

Article Title: Various distribution of BMPs in different periosteal layers contributing to inconsistent osteoinductivity of DBM-based products

doi: 10.1038/s41598-026-35269-z

Figure Lengend Snippet: The trend of BMP-7 distributed in DBM and cortical particles was similar with BMP-2.

Article Snippet: Sandwich enzyme-linked immunosorbent assay (ELISA) was used to assess the amounts of BMP with Commercially available quantikine kits purchased from MULTI SCIENCES (LIANKE) BIOTECH, CO, LTD (human BMP-2 ELISA Kit, sensitivity, 1.73 pg/ml; BMP-7 ELISA Kit, sensitivity, 2.06 pg/ml).

Techniques:

Journal: Scientific Reports

Article Title: Various distribution of BMPs in different periosteal layers contributing to inconsistent osteoinductivity of DBM-based products

doi: 10.1038/s41598-026-35269-z

Figure Lengend Snippet: Through the linear regression analysis, the content of BMP-7 varied linearly with the content of BMP-2 (GuHCl vs. collagenase, R2 = 0.945 vs. R2 = 0.818, respectively).

Article Snippet: Sandwich enzyme-linked immunosorbent assay (ELISA) was used to assess the amounts of BMP with Commercially available quantikine kits purchased from MULTI SCIENCES (LIANKE) BIOTECH, CO, LTD (human BMP-2 ELISA Kit, sensitivity, 1.73 pg/ml; BMP-7 ELISA Kit, sensitivity, 2.06 pg/ml).

Techniques:

Journal: Cell Reports Methods

Article Title: An orthogonal CRISPR/Cpf1 platform for precise spatiotemporal gene regulation and osteoporotic fracture repair

doi: 10.1016/j.crmeth.2025.101299

Figure Lengend Snippet: Multiplex orthogonal gene regulation and programmable editing by OREC with therapeutic application in osteoblasts (A) Experimental illustration of orthogonal activation and repression of ASCL1 and RAB7A gene expression in HEK293T cells using distinct chemical and light inductions. (B) RT-qPCR time course analysis demonstrating orthogonal and reversible control of ASCL1 activation and RAB7A repression in response to alternating Dox and light stimuli. Data represent mean ± SEM ( n = 3). vs. RAB7A baseline (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001); vs. RAB7A 36 h ( $ p < 0.05, $$$ p < 0.001); vs. ASCL1 baseline ( && p < 0.01, &&&& p < 0.0001); vs. ASCL1 48 h ( ## p < 0.01, #### p < 0.0001). (C) Schematic diagram illustrating crRNA guide-length-dependent transcriptional repression and HDR-based gene editing using OREC c system. (D) Transcriptional repression of RAB7A and FZD1 genes following 72 h doxycycline treatment using 15 bp crRNA guides. Data represent mean ± SEM ( n = 3) (∗∗∗∗ p < 0.0001). (E) Luciferase activity analysis following HDR-mediated luciferase knockin using 20 bp crRNA guides. Data represent mean ± SEM ( n = 3) (∗∗∗∗ p < 0.0001). (F) Flow cytometry analysis showing percentage of GFP-positive cells following HDR-mediated GFP knockin. Data represent mean ± SEM ( n = 3) (∗∗∗∗ p < 0.0001). (G) Schematic representation illustrating simultaneous orthogonal activation of Bmp2 and repression of Dkk1 expression in MC3T3-E1 cells. (H) RT-qPCR analysis demonstrating significantly enhanced Bmp2 activation and Dkk1 repression under simultaneous orthogonal induction compared with single-gene regulatory conditions. Data are presented as mean ± SEM ( n = 3). ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (I) Representative images of alizarin red S (ARS) and alkaline phosphatase (ALP) staining showing significantly enhanced osteogenic differentiation following orthogonal OREC induction compared to single-gene controls. Color code for experimental conditions: white: empty vector control; gray: OREC o construct only; crosshatched: OREC c construct only; red: OREC o + Light (optogenetic activation); green: OREC c + Dox (chemogenetic activation) (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (J) RT-qPCR analysis demonstrating synergistic upregulation of osteogenic marker genes ( Alp , Bglap , and Sp7 ) upon simultaneous Bmp2 activation and Dkk1 repression. Data represent mean ± SEM ( n = 3) (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (K) The CCK-8 assay quantifies viable cells via dehydrogenase-mediated formazan formation measured at 450 nm, showing that OREC treatments do not affect cell viability in MC3T3-E1 cells. Data shown as mean ± SEM ( n = 3). See also and .

Article Snippet: Recombinant BMP2 protein , MedChemExpress , Cat# HY-P7006.

Techniques: Multiplex Assay, Activation Assay, Gene Expression, Quantitative RT-PCR, Control, Luciferase, Activity Assay, Knock-In, Flow Cytometry, Expressing, Staining, Plasmid Preparation, Construct, Marker, CCK-8 Assay

Journal: Cell Reports Methods

Article Title: An orthogonal CRISPR/Cpf1 platform for precise spatiotemporal gene regulation and osteoporotic fracture repair

doi: 10.1016/j.crmeth.2025.101299

Figure Lengend Snippet: AAV-delivered OREC system design and in vivo orthogonal gene regulation in fracture healing (A) OREC o system comprises two vectors encoding dAsCpf1-Activer (upper) and TetR-CIBN-P2A-CRY2-VP16 3 with Bmp2 -targeting crRNA (lower). (B) OREC c system consists of vectors encoding dLbCpf1-Repr (upper) and another vector expressing rtTA-Advanced with Dkk1 -targeting crRNA (lower). See also . All constructs designed within AAV packaging constraints. (C) Experimental setup showing AAV intratibial injection procedure with custom LED patch device and power supply system for localized light delivery. (D) Experimental timeline: AAV intratibial injection 14 days before fracture (day 14), tibial fracture induction (day 0), and blue light treatment (470 nm, twice daily for 30 min) beginning one day post-fracture (day 1) through endpoint (day 42). (E and F) AAV2/9 vector constructs for bioluminescence analysis of OREC activity in vivo . (G) Experimental timeline showing AAV intratibial injection (day 0) followed by 14-day treatment with light stimulation (470 nm, 30 min twice daily) or oral doxycycline (30 mg/kg daily) prior to bioluminescence imaging. (H) Representative bioluminescence images demonstrating inducible luciferase expression at tibial sites by OREC with no detectable signal in distant tissues. (I) RT-qPCR analyses of Bmp2 and Dkk1 expression levels in orthogonally treated fracture sites at days 7 and 21 post-fracture. Data represent mean ± SEM ( n = 6 mice/group) (∗∗ p < 0.01, ∗∗∗∗ p < 0.0001). (J) Analysis of Bmp2 and Dkk1 expression relative to Gapdh using the 2 −ΔCt method in fracture callus tissue 7 days after light or doxycycline induction in OREC-transduced mice. Data represent mean ± SEM ( n = 6 mice per group). (K and L) RT-qPCR analysis of Bmp2 (K) and Dkk1 (L) expression in fracture callus tissue from mice exposed to inducers without OREC constructs. Data represent mean ± SEM ( n = 3 mice per group). See also and .

Article Snippet: Recombinant BMP2 protein , MedChemExpress , Cat# HY-P7006.

Techniques: In Vivo, Plasmid Preparation, Expressing, Construct, Injection, Activity Assay, Imaging, Luciferase, Quantitative RT-PCR

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Preparation and in vitro sustained release performance of thermosensitive gel microsphere composite loaded with Anti-Tuberculosis drug PaMZ/BMP-2

doi: 10.1186/s13018-025-06595-1

Figure Lengend Snippet: Flowchart of PaMZ/BMP-2 microsphere preparation process. The outer phase was prepared by dissolving 0.4 g of polyvinyl alcohol in 20 mL of ultrapure water with stirring at 70 °C until completely dissolved, then slowly cooled to obtain a 2% PVA solution. The inner phase was formed by mixing the inner aqueous phase—comprising 4 mg Moxifloxacin, 15 mg Pyrazinamide, and 1 μg BMP-2 dissolved in 1 mL ultrapure water via 5-min ultrasonication—with the oil phase, which contained 1 mg Pretomanid and 46.67 mg PLGA dissolved in 2.5 mL dichloromethane by 3-min ultrasonication. The two phases were combined by vortex mixing followed by 5 min ultrasonication to form the inner phase liquid. Finally, the PaMZ/BMP-2 thermosensitive gel microsphere composite was constructed using microfluidic technology by combining the inner and outer phases, ensuring high drug loading and controlled release

Article Snippet: Internal aqueous phase (M/Z/BMP-2 solution) 10 μg of BMP-2 (HY-P7006, MedChemExpress) was dissolved in 10 μL of acetic acid (HAc) as a stock solution.

Techniques: Construct

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Preparation and in vitro sustained release performance of thermosensitive gel microsphere composite loaded with Anti-Tuberculosis drug PaMZ/BMP-2

doi: 10.1186/s13018-025-06595-1

Figure Lengend Snippet: Morphological characterization diagram of PaMZ/BMP-2 microspheres. A Shows the macroscopic appearance of the microspheres observed with the naked eye. B Presents the morphology of the microspheres captured by a computer terminal during the preparation process. C Displays a 400 × magnified image under an optical microscope. D Shows the surface morphology of drug-loaded microspheres after drying and gold sputtering, as observed by field emission scanning electron microscopy

Article Snippet: Internal aqueous phase (M/Z/BMP-2 solution) 10 μg of BMP-2 (HY-P7006, MedChemExpress) was dissolved in 10 μL of acetic acid (HAc) as a stock solution.

Techniques: Microscopy, Electron Microscopy

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Preparation and in vitro sustained release performance of thermosensitive gel microsphere composite loaded with Anti-Tuberculosis drug PaMZ/BMP-2

doi: 10.1186/s13018-025-06595-1

Figure Lengend Snippet: Particle size distribution map of PaMZ/BMP-2 microspheres

Article Snippet: Internal aqueous phase (M/Z/BMP-2 solution) 10 μg of BMP-2 (HY-P7006, MedChemExpress) was dissolved in 10 μL of acetic acid (HAc) as a stock solution.

Techniques:

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Preparation and in vitro sustained release performance of thermosensitive gel microsphere composite loaded with Anti-Tuberculosis drug PaMZ/BMP-2

doi: 10.1186/s13018-025-06595-1

Figure Lengend Snippet: X-ray diffraction intensity—diffraction Angle curve of PaMZ/BMP-2 microspheres

Article Snippet: Internal aqueous phase (M/Z/BMP-2 solution) 10 μg of BMP-2 (HY-P7006, MedChemExpress) was dissolved in 10 μL of acetic acid (HAc) as a stock solution.

Techniques: