Development of a decellularized porcine bone graft by supercritical carbon dioxide extraction technology for bone regeneration

Abstract

A series of novel decellularized porcine collagen bone graft (DPB) materials in a variety of shapes and sizes were developed by the supercritical carbon dioxide (SCCO2) extraction technique. The complete decellularization of DPB was confirmed by hematoxylin and eosin staining, 4,6‐diamidino‐2‐phenylindole (DAPI) staining, and residual DNA analysis. The native intact collagen remained in the DPB after the SCCO2 process was confirmed by Masson trichrome staining. The physicochemical characteristics of DPB were investigated by scanning electron microscopy and x‐ray diffraction. The cytotoxicity and biocompatibility tests according to ISO10993 and its efficacy for bone regeneration in osteochondral defects in rabbits were evaluated. The rabbit pyrogen test confirmed DPB was non‐toxic. In vitro and in vivo biocompatibility tests of the DPB did not show any toxic or mutagenic effects. The bone regeneration potential of the DPB presented no significant histological differences compared to commercially available deproteinized bovine bone. In conclusion, DPB produced by SCCO2 exhibited similar chemical characteristics to human bone, no toxicity, good biocompatibility, and enhanced bone regeneration in rabbits comparable to that of deproteinized bovine bone. Results from this study could shed light on the potential application of the SCCO2 extraction technique to generate a native decellularized scaffold for bone tissue regeneration in human clinical trials.

Reconstruction of the orbital floor using supercritical CO2 decellularized porcine bone graft

“…It performed as an excellent bone substitute that can regenerate the bone void. ABCcolla ® Collagen Bone Graft increased new bone formation in void sites, indicating good potential for osteoconductivity 4 .…”

“…The scCO 2 process produced ABCcolla ® Collagen Bone Graft does not alter the native structure of bone. The various sizes of porous bone structure maintained after scCO 2 process are essential in angiogenesis and bone growth and bone reorganization in and around the graft material [4,22]. ABCcolla ® Collagen Bone Graft derived from the scCO 2 extraction technology has been proved to be pyrogen-free.…”

“…ABCcolla ® Collagen Bone Graft was manufactured using supercritical carbon dioxide technology and characterized following Chen et al 4 , 13 , 14 .…”

“…Orbital fracture is usually the result of a traumatic accident and high-energy injuries lead to impaired alignment and integrity of facial bones, with the symptoms of periorbital edema, tissue lacerations, enophthalmos and diplopia [1][2][3][4][5][6]. Thus, restoring symmetry of orbital walls is critical to improving clinical outcomes [1-3, 5, 6].…”

“…ABCcolla ® Collagen Bone Graft, a decellularized porcine bone matrix was used in the reconstruction of the orbital floor. It is a porcine cortical bone matrix processed by supercritical carbon dioxide (scCO2) extraction technology to remove cells, fats, hydrocarbons and non-collagenous proteins while preserving the collagen and bone matrix scaffold structure intact 4 , 13 , 14 . scCO 2 extraction technology-mediated decellularization process improved the regenerative nature of the xenogenic graft 15 .…”