J Cosmet Med.  2019 Dec;3(2):86-93. 10.25056/JCM.2019.3.2.86.

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

• Chung S
• Kwon H
• Kim NP

• KMID: 2561076
• DOI: http://doi.org/10.25056/JCM.2019.3.2.86

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.

Keyword

enzyme decellularization; extracellular matrix; regeneration therapeutics; supercritical extraction process