In vitro and in vivo detection of microbial gene expression in bioactivated scaffolds seeded with cyanobacteria

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dc.contributor.authorLeibrock, L. B.
dc.contributor.authorHofmann, D. M.
dc.contributor.authorFuchs, B.
dc.contributor.authorBirt, A.
dc.contributor.authorReinholz, M.
dc.contributor.authorGuertler, A.
dc.contributor.authorFrank, K.
dc.contributor.authorGiunta, R. E.
dc.contributor.authorEgaña Erazo Jose Tomas
dc.contributor.authorNickelsen, J.
dc.contributor.authorSchenck, T. L.
dc.contributor.authorMoellhoff, N.
dc.date.accessioned2024-01-26T12:03:06Z
dc.date.available2024-01-26T12:03:06Z
dc.date.issued2022
dc.description.abstractDermal replacement materials bioactivated with cyanobacteria have shown promising potential for wound regeneration. To date, extraction of cyanobacteria RNA from seeded scaffolds has not been described. The aim of this study was to develop a method to isolate total RNA from bioactivated scaffolds and to propose a new approach in determining living bacteria based on real-time PCR. Transgenic Synechococcus sp. PCC 7002 (tSyn7002) were seeded in liquid cultures or scaffolds for dermal regeneration in vitro and in vivo for 7 days. RNA was extracted with a 260/280 ratio of >= 2. The small subunit of the 30S ribosome in prokaryotes (16S) and RNAse P protein (rnpA) were validated as reference transcripts for PCR analysis. Gene expression patterns differed in vitro and in vivo. Expression of 16S was significantly upregulated in scaffolds in vitro, as compared to liquid cultures, whilst rnpA expression was comparable. In vivo, both 16S and rnpA showed reduced expression compared to in vitro (16S: in vivo Ct value 13.21 +/- 0.32, in vitro 12.44 +/- 0.42; rnpA in vivo Ct value 19.87 +/- 0.41, in vitro 17.75 +/- 1.41). Overall, the results demonstrate rnpA and 16S expression after 7 days of implantation in vitro and in vivo, proving the presence of living bacteria embedded in scaffolds using qPCR.
dc.format.extent9 páginas
dc.fuente.origenWOS
dc.identifier.doi10.1111/lam.13740
dc.identifier.eissn1472-765X
dc.identifier.issn0266-8254
dc.identifier.urihttps://doi.org/10.1111/lam.13740
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/80962
dc.identifier.wosidWOS:000802944700001
dc.information.autorucInstituto de Ingeniería Biológica y Médica; Egaña Erazo Jose Tomas; 0000-0003-2624-6992; 1018715
dc.issue.numero75
dc.language.isoen
dc.nota.accesoContenido parcial
dc.pagina.final409
dc.pagina.inicio401
dc.publisherWILEY
dc.revistaLetters in Applied Microbiology
dc.rightsacceso restringido
dc.subjectWound healing
dc.subjectChronic wounds
dc.subjectCyanobacteria
dc.subjectPhotosynthesis
dc.subject.ddc610
dc.subject.deweyMedicina y salud
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleIn vitro and in vivo detection of microbial gene expression in bioactivated scaffolds seeded with cyanobacteria
dc.typeartículo
dc.volumen2
sipa.codpersvinculados1018715
sipa.trazabilidadWOS;2022-07-08
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