Supercritical extraction of decellularized extracellular matrix from porcine adipose tissue as regeneration therapeutics

Abstract

Background: Extracellular matrix (ECM) has been broadly applied and shown great promise in medical applications. ECM products should be used after decellularization and purification. Supercritical carbon dioxide treatment is of particular interest for purifying ECM due to its medical availability and rapid process speed. However, it is not fully researched for treatment of biomaterials for tissue engineering. Therefore, we investigated the optimal conditions of supercritical carbon dioxide processing at different extracting parameters in porcine adipose tissue. Objective:We aimed to identify the optimal supercritical extracting conditions to produce non-cytotoxic and sterile decellularized extracellular matrix (DE-ECM) for regeneration therapeutics. Methods:The three-day dual treatment including enzymatic decellularization and supercritical fluid extraction of pork adipose tissue was performed. Two protocols using different extracting parameters were applied to evaluate the influence of extracting pressure and temperature on the extraction yield, DNA concentration, and remaining collagen in product. Results:Yield rate increased when high temperature or pressure was applied and pre-enzyme treatment had higher yield rate percent than pre-supercritical processing. Nearly 90% DNA was removed from the pre-enzyme sample when extracted at 3.04×10 7 Pa and 30°C±5°C. The pre-enzyme process had efficient extracting ability at each temperature and pressure and the remaining collagen steadily decreased with increase in extracting pressure and temperature. At the lowest temperature (20°C±5°C) and pressure (1.01×10 7 Pa), remaining collagen was 75.74%±1.83%. Supercritical extraction technology can produce DE-ECM eliminating DNA content efficiently and the remaining proper collagen amount successfully. Conclusion:This study evaluated the feasibility of utilizing supercritical extraction technology in bio-materials and was proven to be is successful. Through controlling the extracting pressure and temperature, this technology has a potential for DE-ECM mass production, which can be useful as tissue regeneration therapeutics as well new drug delivery paradigm.

Supercritical Fluid‐Based Decellularization Technologies for Regenerative Medicine Applications

“…Previous studies suggest that CO 2 solvent pressure between 10 and 30 Mpa displayed limited tissue deformation with effective DNA elimination. [ 14,42–44 ] In particular, it is important to understand the relationship between pressure and temperature in controlling the extraction process of a supercritical fluid. The density, viscosity, and diffusion power, which are the physical components that affect the extraction process yield, can be adjusted by controlling the pressure and temperature.…”

“…As a result of using supercritical fluid after tissue pretreatment with enzymes, the yield rate of the decellularized matrix in the enzyme pretreated group was higher than that of the supercritical fluid only‐treated group, and the decellularization efficiency was increased with the extraction pressure. [ 43 ]…”

Decellularization of bone tissue in a supercritical carbon dioxide environment

“…In paper [17], a comparative analysis of the traditional process of decellularization using detergents and decellularization in SC-CO 2 was carried out. The obtained ECMs were examined using histological examination, scanning electron microscopy (SEM) and DNA and collagen were determined.…”

3D Printing of Meat Following Supercritical Fluid Extraction

“…The SCF process widely applied in the culinary industry so far is as follows. Table 1 illustrates the effective treatment methods used for the extraction of ingredients from various food materials [ 18 , 19 , 20 , 21 ]. The study focuses on providing a platform for scientifically applying the SCF extraction process of meat, followed by 3D printing.…”