Absolute quantification of residual DNA in a new extracellular matrix derived from human reticular dermis (HADM) using real-time TaqMan® MGB-PCR
| Autor: | Massimiliano Bergallo, Ilaria Galliano, Carlotta Castagnoli, Paola Montanari, Daniela Alotto, Angelo Ferravante, Irene Cambieri, Mara Fumagalli, Stefania Casarin, Maurizio Stella |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Acellular Dermis Glycerol NaOH DMEM DNA extraction HADM Taqman PCR Biomaterials Biomedical Engineering Polymerase Chain Reaction law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Dermis law medicine TaqMan Humans 030212 general & internal medicine Polymerase chain reaction Decellularization General Medicine DNA Extracellular Matrix 030104 developmental biology medicine.anatomical_structure chemistry Reticular Dermis Biomedical engineering |
| Zdroj: | Bio-medical materials and engineering. 29(1) |
| ISSN: | 1878-3619 |
| Popis: | Background The development of dermal scaffolds is of major interest in reconstructive surgery. Human Acellular Dermal Matrices (HADMs) provides biomechanical support and elicits new tissue formation. The use of allograft dermis is limited by its immunogenic characteristics. Our research group has focused on the use of human alloplastic glycerolized reticular dermis. Objective The dermal grafts were subjected to two different decellularization protocols in parallel, in order to compare the efficacy in the elimination of residual DNA. Methods It was compared the incubation of the dermis in NaOH (0.06 N) and in the standard culture medium "Dulbecco Modified Eagle Medium" (DMEM). The samples were incubated in the specific medium for 8 weeks. The newly developed real-time TaqMan® MGB-PCR assay was applied for both the detection and absolute quantification of residual DNA. Results It was observed that the level of residual DNA decreased until time T3 and remained constant until time T8. Moreover, there was no statistical difference between treatment with DMEM or NaOH 0.06 N as to the amount of residual DNA. Conclusions Decellularization methods, DMEM or NaOH 0.06 N do not affect DNA recovery. The proposed approach offers an alternative method to quantify residual DNA in HADM samples. |
| Databáze: | OpenAIRE |
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