Evaluation of freeze‐drying and cryopreservation protocols for long‐term storage of biomaterials based on decellularized intestine

Abstract

We evaluated the efficiency of several protocols to preserve the main components of decellularized tissue scaffolds for delayed use. Decellularized rat intestine scaffolds were generated by using SDS and triton X-100 and preserved for 3 months subjected to eight freeze-drying (F1 to F8) and 14 cryopreservation protocols (C1 to C14). Morphological analysis showed that cryopreservation tended to preserve the tissue morphostructure more efficiently than freeze-drying. Histological analysis showed that the content of proteoglycans and glycoproteins was efficiently preserved by most methods. The protocols that most efficiently preserved collagen fibers were those using trehalose and saccharose for freeze-drying (F2, F3, and F7 protocols) and DMSO, albumin, and saccharose (C3, C5, C6, C12) for cryopreservation. Most freeze-drying protocols and cryopreservation protocols with DMSO, albumin, and maltose (C6, C7, C13, and C14) efficiently preserved reticular fibers. For the elastic fibers, freeze-drying methods with trehalose and maltose (F2, F4, F6, and F8) properly preserved these fibers, with the results of most cryopreservation methods comparable to controls. These results suggest that freeze-drying using 0.1M trehalose and cryopreservation in the presence of 8% DMSO and 4.6% albumin are more efficient than other protocols in preserving the scaffold morphostructure and histological composition. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 488-500, 2018.

Long-term preservation effects on biological properties of acellular placental sponge patches

“…The preservation condition is also very critical, which affects remarkably the quality of the ECM based scaffolds. It is shown that preservation methods and storage time can remarkably influence the biomechanical and biophysical properties of the decellularized tissues [14][15][16].The decellularized tissues can be stored through the different preservation methods until experimental or clinical uses. Freeze-drying and cryopreservation are two common preservation methods for long-time preservation of tissue-derived tissue engineering products [14,15].…”

Influence of Storage Conditions on Decellularized Porcine Conjunctiva

“…For completeness, it is worth mentioning that there are other storage forms described for acellular tissues not investigated in the current study, such as vitrification [ 28 , 47 ] or freeze drying [ 28 , 48 , 49 ]. Freeze drying is of interest because it allows storage at room temperature and thus, saves storage costs, enables easy transportation and does not require potentially toxic protective agents [ 24 ].…”

Development of a large‐volume human‐derived adipose acellular allogenic flap by perfusion decellularization

“…Future research should also define the ideal methods for long‐term preservation of tissues before and after decellularization. Extensive and successful clinical experience with commercially available decellularized small‐volume tissues suggests that both cryopreservation and freeze‐drying might provide an effective solution, applicable to large‐volume tissues (e.g., flaps) and organs …”

“…Extensive and successful clinical experience with commercially available decellularized small-volume tissues suggests that both cryopreservation and freezedrying might provide an effective solution, applicable to large-volume tissues (e.g., flaps) and organs. [42][43][44][45] In conclusion, we here report for the first time the successful creation of a human-derived, large-volume AAF. We envision that this study may establish the preliminary basis for the development of off-the-shelf AAFs that could disrupt current standards of care in reconstructive surgery and benefit a large number of patients in the future.…”