Protocols for accelerated production and purification of collagen scaffold and atelocollagen from animal tissues

Traditional purification of atelocollagen involves a harsh extraction process with environment-polluting chemicals and costly and/or time-consuming procedures which include salting-out, alkali, acid and enzymatic treatment and ion-exchange chromatography. The atelocollagen market is growing exponentially, with demand in the skincare industry and for various medical applications. As a result, there is an urgent need for an eco-friendly production process with minimal manipulation. We developed a novel technique involving supercritical carbon dioxide extraction technology to remove the cells and noncollagenous substances from the porcine hide. Subsequent processes allow the production of several products, including decellularized dermal membrane, high-purity collagen particles and atelocollagen. The advantages of our process are its faster speed and lower environmental impact and its generation of multiple products, including high purity atelocollagen with complete removal of telopeptides. Sliced porcine skin was subjected to a proprietary supercritical carbon dioxide extraction technology for decellularization. The decellularized dermis collagen scaffold was freeze-dried and freeze-milled to granules of 50–200 μm and subjected to enzymatic hydrolysis using pepsin in acidic conditions, then subjected to ultrafiltration for pepsin and telopeptide removal. The atelocollagen solution was filtered through a 0.2-μm filter for sterilization. The acidic atelocollagen solution was subjected to fibrillogenesis by bringing the pH to 7, then centrifuged to obtain the atelocollagen slurry. This slurry was then freeze-dried to obtain atelocollagen dry powder. The atelocollagen solution was characterized using SDS-PAGE.