A decellularized extracellular matrix (ECM) scaffold is a biomaterial made by removing the cells from natural tissue while preserving the surrounding ECM structure. It is the dominant technical theme across the patent portfolio collected in the source archive Bone Glue dossier, appearing in the majority of the eighteen patent entries.
The method
The patents in the dossier describe a recurring sequence of treatments applied to natural tissue — bone, periosteum, tendon-conjunction bone, synchondrosis bone, epiphyseal cartilage combined bone, or intervertebral disc. The reagents named across the set include:
- Protease inhibitors in saline or PBS buffer, to limit enzymatic degradation of the matrix.
- Triton X-100 in PBS, a non-ionic detergent used to lyse cell membranes.
- SDS (sodium dodecyl sulfate) or SLES in PBS, an anionic detergent used to strip residual cellular material.
- DNase I in PBS, to digest residual DNA.
- Freeze–thaw cycles, sometimes with liquid nitrogen, to disrupt cells.
- Ultrasonic treatment, used for decalcification or additional disruption.
- Organic solvents, for degreasing.
- EDTA isotonic solution, in some protocols.
The stated goal is to remove immunogenic allogeneic or xenogeneic cells completely while preserving the ECM's three-dimensional structure and active components.
Representative protocols from the dossier
CN104307045 (decellularized periosteum): rinse three times in aseptic PBS; oscillate 1 hour in 5% PBS with 10 KIU/ml protease inhibitor at 200 rpm; 48 hours in 5% PBS with TritonX-100 at 250 rpm; 48 hours in 10% PBS with SDS at 250 rpm; 12 hours in 1.5 mg/ml PBS with DNAase at 250 rpm.
CN104307044 (total disc acellular material): intervertebral disc of a vertebrate; 4 hours at 150 rpm in 10% PBS with 10 KIU/ml; 48 hours in 4% PBS with TritonX-100; 48 hours in 5% PBS with SDS; 12 hours in 0.5 mg/ml PBS with DNA enzyme; final PBS flush for 1 hour.
WO2023040853 (periosteum-bone complex): cutting and proofing, repeated deionized-water rinsing, liquid-nitrogen freeze–thaw, ultrasonic decalcification, then PBS with protease inhibitor, PBS with Triton X-100, PBS with SLES, PBS with DNase I and Tris-HCl buffer.
Mineralization control
Several patents in the dossier combine decellularization with controlled demineralization:
- CN116328039 targets a calcium mass content of 10–20% after ultrasonic decellularization and demineralization, producing a material with surface charged groups for selective metabolite permeation and surface collagen residues for cell adhesion.
- US11684696 applies gradient demineralization to decellularized bone, expanding porosity and surface collagen exposure, releasing growth factors and improving cell adhesion.
- CN112618797 uses a specific demineralized and decellularized cancellous bone ECM scaffold (SDECM) as a substrate for antibiotics loaded by electrostatic adsorption and chemical crosslinking, released by pH-responsive and degradation-accompanying modes.
Evidence status
These are patent abstracts. The dossier provides no grant status, prosecution outcome or experimental validation for any of them. A patent is a legal claim, not proof of effectiveness — see Patent as Evidence. The dossier does not state that any of these scaffolds is the material sold or reported as Bone-02.
Related
Source notes & attribution
- https://rexresearch.com/XianfengBoneGlue/XianfengBoneGlue.html