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Fulltext Cocktail Protocol for Preparation of Platelet-Rich Fibrin Glue for Autologous Use

Chenna D, Shastry S, Das S

Malays J Med Sci, 2021 Feb;28(1):35-40.
PMID: 33679218 DOI: 10.21315/mjms2021.28.1.5

Background: Biomaterials containing platelets have been used to promote healing of ulcers and burns, as well as in implantology and maxillofacial and plastic surgery to achieve wound healing and tissue repair. Commercial devices to prepare autologous biomaterials involve diverse preparation methods that can have high production costs and low yields. Hence, we designed a protocol for preparation of large amounts of autologous platelet-rich fibrin (PRF) glue using conventional processing techniques for blood components.
Methods: Autologous whole blood collected 72 h before surgery was processed to prepare platelet concentrates and cryoprecipitate. In a closed system, calcium was added to the cryoprecipitate to release autologous thrombin and generate a firm fibrin clot. The fibrin clot, platelets and calcium were then placed in a conical flask in which a PRF glue formed. The protocol was validated through determination of pre- and post-platelet counts and fibrinogen amounts in the product.
Results: Platelets were recovered with 68% efficiency during the preparation. Essentially no platelets or fibrinogen were found in the supernatant of the PRF glue, suggesting that nearly all had been incorporated in a PRF glue having a relatively large (8 cm × 10 cm) size.
Conclusion: The protocol described here is a cost-effective, simple and closed system that can be used to produce large-size PRF glue to promote repair of major surgical defects.

Matched MeSH terms: Thrombin
Fulltext Characterisation of aptamer-anchored poly(EDMA-co-GMA) monolith for high throughput affinity binding

Acquah C, Chan YW, Pan S, Yon LS, Ongkudon CM, Guo H, et al.

Sci Rep, 2019 10 10;9(1):14501.
PMID: 31601836 DOI: 10.1038/s41598-019-50862-1

Immobilisation of aptameric ligands on solid stationary supports for effective binding of target molecules requires understanding of the relationship between aptamer-polymer interactions and the conditions governing the mass transfer of the binding process. Herein, key process parameters affecting the molecular anchoring of a thrombin-binding aptamer (TBA) onto polymethacrylate monolith pore surface, and the binding characteristics of the resulting macroporous aptasensor were investigated. Molecular dynamics (MD) simulations of the TBA-thrombin binding indicated enhanced Guanine 4 (G4) structural stability of TBA upon interaction with thrombin in an ionic environment. Fourier-transform infrared spectroscopy and thermogravimetric analyses were used to characterise the available functional groups and thermo-molecular stability of the immobilised polymer generated with Schiff-base activation and immobilisation scheme. The initial degradation temperature of the polymethacrylate stationary support increased with each step of the Schiff-base process: poly(Ethylene glycol Dimethacrylate-co-Glycidyl methacrylate) or poly(EDMA-co-GMA) [196.0 °C (±1.8)]; poly(EDMA-co-GMA)-Ethylenediamine [235.9 °C (±6.1)]; poly(EDMA-co-GMA)-Ethylenediamine-Glutaraldehyde [255.4 °C (±2.7)]; and aptamer-modified monolith [273.7 °C (±2.5)]. These initial temperature increments reflected in the associated endothermic energies were determined with differential scanning calorimetry. The aptameric ligand density obtained after immobilisation was 480 pmol/μL. Increase in pH and ionic concentration affected the surface charge distribution and the binding characteristics of the aptamer-modified disk-monoliths, resulting in the optimum binding pH and ionic concentration of 8.0 and 5 mM Mg2+, respectively. These results are critical in understanding and setting parametric constraints indispensable to develop and enhance the performance of aptasensors.

Matched MeSH terms: Thrombin

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