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Use of adipose-derived mesenchymal stem cells in keratoconjunctivitis sicca in a canine model.

Villatoro AJ, Fernández V, Claros S, Rico-Llanos GA, Becerra J, Andrades JA - Biomed Res Int (2015)

Bottom Line: Average STT values and all clinical signs showed a statistically significant change (P < 0.001) during the follow-up with reduction in all ocular parameters scored: ocular discharge, conjunctival hyperaemia, and corneal changes, and there were no signs of regression or worsening.Implanted cells were well tolerated and were effective reducing clinical signs of KCS with a sustained effect during the study period.These results could reinforce a good effective solution to be extrapolated to future studies in human.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bioengineering and Tissue Regeneration (LABRET), Department of Cell Biology, Genetics and Physiology, Faculty of Sciences, Biomedical Research Institute of Málaga (IBIMA), University of Málaga, 29071 Málaga, Spain.

ABSTRACT
Keratoconjunctivitis sicca (KCS) or dry eye disease (DED) is an immune-mediated multifactorial disease, with high level of prevalence in humans and dogs. Our aim in this study was to investigate the therapeutic effects of allogeneic adipose-derived mesenchymal stromal cells (Ad-MSCs) implanted around the lacrimal glands in 12 dogs (24 eyes) with KCS, which is refractory to current available treatments. Schirmer tear test (STT) and ocular surface integrity were assessed at 0 (before treatment), 3, 6, and 9 months after treatment. Average STT values and all clinical signs showed a statistically significant change (P < 0.001) during the follow-up with reduction in all ocular parameters scored: ocular discharge, conjunctival hyperaemia, and corneal changes, and there were no signs of regression or worsening. Implanted cells were well tolerated and were effective reducing clinical signs of KCS with a sustained effect during the study period. None of the animals showed systemic or local complications during the study. To our knowledge, this is the first time in literature that implantation of allogeneic Ad-MSCs around lacrimal glands has been found as an effective therapeutic alternative to treat dogs with KCS. These results could reinforce a good effective solution to be extrapolated to future studies in human.

No MeSH data available.


Related in: MedlinePlus

Assessment of trilineage differentiation. (a) Control. Cells maintained in control medium for 21 days. (b-c) Assessment of osteogenic differentiation. (b) Histochemical localization of ALP. Osteoinduced cells formed numerous nodules highly positive for ALP staining. (c) Positive Alizarin Red S staining by day 21 whereas red calcium nodules clearly appeared on the osteoinduced cultures. (d) Assessment of adipogenic differentiation. Positive Oil Red O staining by day 21 confirmed the presence of lipid droplets only in adipogenic-induced cells. (e-f) Assessment of chondrogenic differentiation. Histological sections of pellets after 21 days in the presence or absence of rhTGF-β1. (e) Control pellets incubated without rhTGF-β1. (f) Pellets incubated with rhTGF-β1 clearly displayed improved chondrogenesis with positive toluidine blue staining. Bars 200 μm in (a–d) and 100 μm in (e-f).
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fig3: Assessment of trilineage differentiation. (a) Control. Cells maintained in control medium for 21 days. (b-c) Assessment of osteogenic differentiation. (b) Histochemical localization of ALP. Osteoinduced cells formed numerous nodules highly positive for ALP staining. (c) Positive Alizarin Red S staining by day 21 whereas red calcium nodules clearly appeared on the osteoinduced cultures. (d) Assessment of adipogenic differentiation. Positive Oil Red O staining by day 21 confirmed the presence of lipid droplets only in adipogenic-induced cells. (e-f) Assessment of chondrogenic differentiation. Histological sections of pellets after 21 days in the presence or absence of rhTGF-β1. (e) Control pellets incubated without rhTGF-β1. (f) Pellets incubated with rhTGF-β1 clearly displayed improved chondrogenesis with positive toluidine blue staining. Bars 200 μm in (a–d) and 100 μm in (e-f).

Mentions: Adipogenic differentiation was confirmed by Oil Red O staining. After culturing cells with adipogenic-inducing media for 21 days, red-stained lipid droplets were present in the cytoplasm (Figure 3(d)).


Use of adipose-derived mesenchymal stem cells in keratoconjunctivitis sicca in a canine model.

Villatoro AJ, Fernández V, Claros S, Rico-Llanos GA, Becerra J, Andrades JA - Biomed Res Int (2015)

Assessment of trilineage differentiation. (a) Control. Cells maintained in control medium for 21 days. (b-c) Assessment of osteogenic differentiation. (b) Histochemical localization of ALP. Osteoinduced cells formed numerous nodules highly positive for ALP staining. (c) Positive Alizarin Red S staining by day 21 whereas red calcium nodules clearly appeared on the osteoinduced cultures. (d) Assessment of adipogenic differentiation. Positive Oil Red O staining by day 21 confirmed the presence of lipid droplets only in adipogenic-induced cells. (e-f) Assessment of chondrogenic differentiation. Histological sections of pellets after 21 days in the presence or absence of rhTGF-β1. (e) Control pellets incubated without rhTGF-β1. (f) Pellets incubated with rhTGF-β1 clearly displayed improved chondrogenesis with positive toluidine blue staining. Bars 200 μm in (a–d) and 100 μm in (e-f).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4352730&req=5

fig3: Assessment of trilineage differentiation. (a) Control. Cells maintained in control medium for 21 days. (b-c) Assessment of osteogenic differentiation. (b) Histochemical localization of ALP. Osteoinduced cells formed numerous nodules highly positive for ALP staining. (c) Positive Alizarin Red S staining by day 21 whereas red calcium nodules clearly appeared on the osteoinduced cultures. (d) Assessment of adipogenic differentiation. Positive Oil Red O staining by day 21 confirmed the presence of lipid droplets only in adipogenic-induced cells. (e-f) Assessment of chondrogenic differentiation. Histological sections of pellets after 21 days in the presence or absence of rhTGF-β1. (e) Control pellets incubated without rhTGF-β1. (f) Pellets incubated with rhTGF-β1 clearly displayed improved chondrogenesis with positive toluidine blue staining. Bars 200 μm in (a–d) and 100 μm in (e-f).
Mentions: Adipogenic differentiation was confirmed by Oil Red O staining. After culturing cells with adipogenic-inducing media for 21 days, red-stained lipid droplets were present in the cytoplasm (Figure 3(d)).

Bottom Line: Average STT values and all clinical signs showed a statistically significant change (P < 0.001) during the follow-up with reduction in all ocular parameters scored: ocular discharge, conjunctival hyperaemia, and corneal changes, and there were no signs of regression or worsening.Implanted cells were well tolerated and were effective reducing clinical signs of KCS with a sustained effect during the study period.These results could reinforce a good effective solution to be extrapolated to future studies in human.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bioengineering and Tissue Regeneration (LABRET), Department of Cell Biology, Genetics and Physiology, Faculty of Sciences, Biomedical Research Institute of Málaga (IBIMA), University of Málaga, 29071 Málaga, Spain.

ABSTRACT
Keratoconjunctivitis sicca (KCS) or dry eye disease (DED) is an immune-mediated multifactorial disease, with high level of prevalence in humans and dogs. Our aim in this study was to investigate the therapeutic effects of allogeneic adipose-derived mesenchymal stromal cells (Ad-MSCs) implanted around the lacrimal glands in 12 dogs (24 eyes) with KCS, which is refractory to current available treatments. Schirmer tear test (STT) and ocular surface integrity were assessed at 0 (before treatment), 3, 6, and 9 months after treatment. Average STT values and all clinical signs showed a statistically significant change (P < 0.001) during the follow-up with reduction in all ocular parameters scored: ocular discharge, conjunctival hyperaemia, and corneal changes, and there were no signs of regression or worsening. Implanted cells were well tolerated and were effective reducing clinical signs of KCS with a sustained effect during the study period. None of the animals showed systemic or local complications during the study. To our knowledge, this is the first time in literature that implantation of allogeneic Ad-MSCs around lacrimal glands has been found as an effective therapeutic alternative to treat dogs with KCS. These results could reinforce a good effective solution to be extrapolated to future studies in human.

No MeSH data available.


Related in: MedlinePlus