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Grafting and early expression of growth factors from adipose-derived stem cells transplanted into the cochlea, in a Guinea pig model of acoustic trauma.

Fetoni AR, Lattanzi W, Eramo SL, Barba M, Paciello F, Moriconi C, Rolesi R, Michetti F, Troiani D, Paludetti G - Front Cell Neurosci (2014)

Bottom Line: ASC implantation did not modify auditory function.ASCs migrated from the perilymphatic to the endolymphatic compartment, during the analyzed time course.Immunofluorescence confirmed the increased expression, which appeared to be further strengthened by ASCs' implantation.

View Article: PubMed Central - PubMed

Affiliation: Department of Head and Neck Surgery, Università Cattolica del Sacro Cuore , Rome , Italy.

ABSTRACT
Noise exposure causes damage of multiple cochlear cell types producing permanent hearing loss with important social consequences. In mammals, no regeneration of either damaged hair cells or auditory neurons has been observed and no successful treatment is available to achieve a functional recovery. Loads of evidence indicate adipose-derived stem cells (ASCs) as promising tools in diversified regenerative medicine applications, due to the high degree of plasticity and trophic features. This study was aimed at identifying the path of in vivo cell migration and expression of trophic growth factors, upon ASCs transplantation into the cochlea, following noise-induced injury. ASCs were isolated in primary culture from the adipose tissue of a guinea pig, transduced using a viral vector to express the green fluorescent protein, and implanted into the scala tympani of deafened animals. Auditory function was assessed 3 and 7 days after surgery. The expression of trophic growth factors was comparatively analyzed using real-time PCR in control and noise-injured cochlear tissues. Immunofluorescence was used to assess the in vivo localization and expression of trophic growth factors in ASCs and cochleae, 3 and 7 days following homologous implantation. ASC implantation did not modify auditory function. ASCs migrated from the perilymphatic to the endolymphatic compartment, during the analyzed time course. Upon noise exposure, the expression of chemokine ligands and receptors related to the PDGF, VEGF, and TGFbeta pathways, increased in the cochlear tissues, possibly guiding in vivo cell migration. Immunofluorescence confirmed the increased expression, which appeared to be further strengthened by ASCs' implantation. These results indicated that ASCs are able to migrate at the site of tissue damage and express trophic factors, upon intracochlear implantation, providing an original proof of principle, which could pave the way for further developments of ASC-based treatments of deafness.

No MeSH data available.


Related in: MedlinePlus

Differential gene expression across different cochlear regions. Graphs display the results obtained using quantitative real-time PCR to analyze the expression of genes in the three microdissected regions of the cochleae of control animals (A–B) and noise-exposed animals (C–D). In (E), real-time PCR data are shown from the paired analysis of control versus noise-exposed animals, in each tested region: stria, stria vascularis; corti, organ of Corti; ganglion, spiral ganglion. In each panel, relative quantity (RQ) refers to the mean levels of gene expression across replicates, calculated using the ΔΔCt method (see text for details).
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Figure 3: Differential gene expression across different cochlear regions. Graphs display the results obtained using quantitative real-time PCR to analyze the expression of genes in the three microdissected regions of the cochleae of control animals (A–B) and noise-exposed animals (C–D). In (E), real-time PCR data are shown from the paired analysis of control versus noise-exposed animals, in each tested region: stria, stria vascularis; corti, organ of Corti; ganglion, spiral ganglion. In each panel, relative quantity (RQ) refers to the mean levels of gene expression across replicates, calculated using the ΔΔCt method (see text for details).

Mentions: First, we have analyzed comparatively the relative gene expression levels across the three cochlear regions, to explain tissue-specific profiles possibly involved in driving cell migration path, in control cochleae (Figures 3A,B), and in noise-exposed cochleae (Figures 3C,D). This revealed that the basal expression of Tgfβ, Ccl2, Ccl5/Rantes (Figure 3A), Vegf-A, and Vegfr2 (Figure 3B) genes did not vary significantly across the three microdissected regions, in control animals. Conversely, the expression of Pdgfa, Pdgfr (Figure 3A), and Vegfr3 (Figure 3B) was significantly reduced in the organ of Corti and in the stria vascularis, compared to the spiral ganglion. Finally, Vegfr1 expression was significantly higher in the organ of Corti, compared to the other regions (Figure 3B).


Grafting and early expression of growth factors from adipose-derived stem cells transplanted into the cochlea, in a Guinea pig model of acoustic trauma.

Fetoni AR, Lattanzi W, Eramo SL, Barba M, Paciello F, Moriconi C, Rolesi R, Michetti F, Troiani D, Paludetti G - Front Cell Neurosci (2014)

Differential gene expression across different cochlear regions. Graphs display the results obtained using quantitative real-time PCR to analyze the expression of genes in the three microdissected regions of the cochleae of control animals (A–B) and noise-exposed animals (C–D). In (E), real-time PCR data are shown from the paired analysis of control versus noise-exposed animals, in each tested region: stria, stria vascularis; corti, organ of Corti; ganglion, spiral ganglion. In each panel, relative quantity (RQ) refers to the mean levels of gene expression across replicates, calculated using the ΔΔCt method (see text for details).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4202717&req=5

Figure 3: Differential gene expression across different cochlear regions. Graphs display the results obtained using quantitative real-time PCR to analyze the expression of genes in the three microdissected regions of the cochleae of control animals (A–B) and noise-exposed animals (C–D). In (E), real-time PCR data are shown from the paired analysis of control versus noise-exposed animals, in each tested region: stria, stria vascularis; corti, organ of Corti; ganglion, spiral ganglion. In each panel, relative quantity (RQ) refers to the mean levels of gene expression across replicates, calculated using the ΔΔCt method (see text for details).
Mentions: First, we have analyzed comparatively the relative gene expression levels across the three cochlear regions, to explain tissue-specific profiles possibly involved in driving cell migration path, in control cochleae (Figures 3A,B), and in noise-exposed cochleae (Figures 3C,D). This revealed that the basal expression of Tgfβ, Ccl2, Ccl5/Rantes (Figure 3A), Vegf-A, and Vegfr2 (Figure 3B) genes did not vary significantly across the three microdissected regions, in control animals. Conversely, the expression of Pdgfa, Pdgfr (Figure 3A), and Vegfr3 (Figure 3B) was significantly reduced in the organ of Corti and in the stria vascularis, compared to the spiral ganglion. Finally, Vegfr1 expression was significantly higher in the organ of Corti, compared to the other regions (Figure 3B).

Bottom Line: ASC implantation did not modify auditory function.ASCs migrated from the perilymphatic to the endolymphatic compartment, during the analyzed time course.Immunofluorescence confirmed the increased expression, which appeared to be further strengthened by ASCs' implantation.

View Article: PubMed Central - PubMed

Affiliation: Department of Head and Neck Surgery, Università Cattolica del Sacro Cuore , Rome , Italy.

ABSTRACT
Noise exposure causes damage of multiple cochlear cell types producing permanent hearing loss with important social consequences. In mammals, no regeneration of either damaged hair cells or auditory neurons has been observed and no successful treatment is available to achieve a functional recovery. Loads of evidence indicate adipose-derived stem cells (ASCs) as promising tools in diversified regenerative medicine applications, due to the high degree of plasticity and trophic features. This study was aimed at identifying the path of in vivo cell migration and expression of trophic growth factors, upon ASCs transplantation into the cochlea, following noise-induced injury. ASCs were isolated in primary culture from the adipose tissue of a guinea pig, transduced using a viral vector to express the green fluorescent protein, and implanted into the scala tympani of deafened animals. Auditory function was assessed 3 and 7 days after surgery. The expression of trophic growth factors was comparatively analyzed using real-time PCR in control and noise-injured cochlear tissues. Immunofluorescence was used to assess the in vivo localization and expression of trophic growth factors in ASCs and cochleae, 3 and 7 days following homologous implantation. ASC implantation did not modify auditory function. ASCs migrated from the perilymphatic to the endolymphatic compartment, during the analyzed time course. Upon noise exposure, the expression of chemokine ligands and receptors related to the PDGF, VEGF, and TGFbeta pathways, increased in the cochlear tissues, possibly guiding in vivo cell migration. Immunofluorescence confirmed the increased expression, which appeared to be further strengthened by ASCs' implantation. These results indicated that ASCs are able to migrate at the site of tissue damage and express trophic factors, upon intracochlear implantation, providing an original proof of principle, which could pave the way for further developments of ASC-based treatments of deafness.

No MeSH data available.


Related in: MedlinePlus