Ultrarapid Purification of Collagen Type I for Tissue Engineering Applications

Abstract

Acid-solubilized collagen type I (COL1) can form highly organized, three-dimensional scaffolds for a wide variety of bioengineering and cell culture applications. A rapid COL1 isolation method would be a valuable tool for both basic and translational researchers because conventional techniques require days or weeks to complete, typically use nonhuman animal tissues as a source material, and do not efficiently purify autologous COL1 from small samples. Here, we describe a 3-h method to isolate COL1 from rabbit, lamb, and human skin in sufficient quantities for fabrication of autologously derived tissues by using a rapid agitation technique and incorporating centrifugal filtration units into a traditional isolation procedure. We demonstrate that the purified product is comparable to traditional preparations using polyacrylamide gel electrophoresis, transmission electron microscopy, and collagen content assays. In addition, our COL1 is able to support myogenic cell growth and direct orientation of these cells in vitro. Importantly, this ultrarapid COL1 isolation procedure increases the feasibility of autologous COL1 use in humans as well as overall safety for clinical patients.

Ectopic osteogenesis by type I collagen loaded with a novel synthesized PTH‐related peptide‐1 in vivo

“…In mammals, collagen is widely available, such as in skin, tendons, bones, and teeth. Rat‐tail Col‐I has a relatively weak immunogenic reaction, and its degradation products have no cytotoxic side effects; collagen gel has good plasticity and encapsulation advantages and has been widely used in many biological materials (Lees, ; Pacak, Powers, & Cowan, ; Rajan, Habermehl, Coté, Doillon, & Mantovani, ). Good osteoinduction and bone conduction ability are important evaluation indexes for bone tissue engineering scaffolds.…”

An Improved Method for the Preparation of Type I Collagen From Skin

“…Conversely, COL1 can be purified from whole rat tails or isolated rat tail tendons using a slightly modified preparation that includes lysing matrix beads. This has been previously described in detail 8 .…”

“…This method is simple to perform using standard buffers and equipment found in many laboratories. By employing high-speed agitation, our protocol reduces the time necessary to isolate soluble, dermal COL1 from approximately 10 days to less than 3 hr 8 . Importantly, this method can be easily performed in the clinical setting in order to prepare autologouslyderived COL1 for use in patients during a single set of procedures.…”

Production of recombinant collagen: state of the art and challenges

“…As the whole extraction process can take up to a month [30], optimisation is necessary to reduce time spent. To achieve a shorter extraction procedure, Pacak et al [29] shortened the time needed for each step significantly. They also replaced the lengthy dialysis process with centrifugation in a 100 k molecular weight cut‐off (MWCO) centrifugal filter tube, saving several days.…”