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Mesenchymal stromal cells for cutaneous wound healing in a rabbit model: pre-clinical study applicable in the pediatric surgical setting.

Pelizzo G, Avanzini MA, Icaro Cornaglia A, Osti M, Romano P, Avolio L, Maccario R, Dominici M, De Silvestri A, Andreatta E, Costanzo F, Mantelli M, Ingo D, Piccinno S, Calcaterra V - J Transl Med (2015)

Bottom Line: Rabbit ASCs were isolated and expanded in vitro with relative abundance, cells expressed typical surface markers (CD49e, CD90 and CD29).Improved re-epithelization, reduced inflammatory infiltration and increased collagen deposition were observed in biopsies from wounds treated with ASCs, with the best result in the autologous setting.Pre-clinical studies are however necessary to validate the best skin regeneration technique, which could be used in pediatric surgical translational research.

View Article: PubMed Central - PubMed

Affiliation: Pediatric Surgery Unit, Fondazione IRCCS Policlinico San Matteo and University of Pavia, 27100, Pavia, Italy. g.pelizzo@smatteo.pv.it.

ABSTRACT

Objective: Mesenchymal stromal cells (MSCs) expanded in vitro have been proposed as a potential therapy for congenital or acquired skin defects in pediatrics. The aim of this pre-clinical study was to investigate the effects of intradermal injections of MSC in experimental cutaneous wound repair comparing allogeneic and autologous adipose stem cells (ASCs) and autologous bone marrow-mesenchymal stromal cells (BM-MSCs).

Methods: Mesenchymal stromal cells were in vitro expanded from adipose and BM tissues of young female New Zealand rabbits. MSCs were characterized for plastic adhesion, surface markers, proliferation and differentiation capacity. When an adequate number of cells (ASCs 10 × 10(6) and BM-MSCs 3 × 10(6), because of their low rate of proliferation) was reached, two skin wounds were surgically induced in each animal. The first was topically treated with cell infusions, the second was used as a control. The intradermal inoculation included autologous or allogeneic ASCs or autologous BM-MSCs. For histological examination, animals were sacrificed and wounds were harvested after 11 and 21 days of treatment.

Results: Rabbit ASCs were isolated and expanded in vitro with relative abundance, cells expressed typical surface markers (CD49e, CD90 and CD29). Topically, ASC inoculation provided more rapid wound healing than BM-MSCs and controls. Improved re-epithelization, reduced inflammatory infiltration and increased collagen deposition were observed in biopsies from wounds treated with ASCs, with the best result in the autologous setting. ASCs also improved restoration of skin architecture during wound healing.

Conclusion: The use of ASCs may offer a promising solution to treat extended wounds. Pre-clinical studies are however necessary to validate the best skin regeneration technique, which could be used in pediatric surgical translational research.

No MeSH data available.


Related in: MedlinePlus

Characterization of rabbit MSCs. A Morphology of bone marrow and adipose-derived mesenchymal stromal cells obtained from one rabbit. MSCs from both sources display the characteristic spindle-shaped morphology. Magnification ×4. B Immunophenotype of culture-expanded ASCs obtained from one representative rabbit. ASCs were positive for CD90, CD29 and CD49e and negative for CD45 and CD10, as reported [20]. The immunophenotype of BM-MSCs was superimposable. C Osteogenic and adipogenic differentiation capacity of BM-MSCs and ASC. Differentiation into osteoblasts was demonstrated by the histological detection of Alkaline phosphatase activity (a) and calcium depositions positive for Alizarin Red (b); magnification ×20. Differentiation into adipocytes was revealed by the formation of lipid droplets stained with Oil Red O (c); magnification ×20. Non differentiated cells (negative ctrl) are reported.
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Fig2: Characterization of rabbit MSCs. A Morphology of bone marrow and adipose-derived mesenchymal stromal cells obtained from one rabbit. MSCs from both sources display the characteristic spindle-shaped morphology. Magnification ×4. B Immunophenotype of culture-expanded ASCs obtained from one representative rabbit. ASCs were positive for CD90, CD29 and CD49e and negative for CD45 and CD10, as reported [20]. The immunophenotype of BM-MSCs was superimposable. C Osteogenic and adipogenic differentiation capacity of BM-MSCs and ASC. Differentiation into osteoblasts was demonstrated by the histological detection of Alkaline phosphatase activity (a) and calcium depositions positive for Alizarin Red (b); magnification ×20. Differentiation into adipocytes was revealed by the formation of lipid droplets stained with Oil Red O (c); magnification ×20. Non differentiated cells (negative ctrl) are reported.

Mentions: ASCs and BM-MSCs showed the typical spindle shape morphology (Figure 2, Panel A) and they resulted positive for CD49e, CD90 and CD29, while they were negative for CD45 and CD10 (Figure 2, Panel B for in vitro-expanded ASCs) as reported by Piccinno et al. [21].Figure 2


Mesenchymal stromal cells for cutaneous wound healing in a rabbit model: pre-clinical study applicable in the pediatric surgical setting.

Pelizzo G, Avanzini MA, Icaro Cornaglia A, Osti M, Romano P, Avolio L, Maccario R, Dominici M, De Silvestri A, Andreatta E, Costanzo F, Mantelli M, Ingo D, Piccinno S, Calcaterra V - J Transl Med (2015)

Characterization of rabbit MSCs. A Morphology of bone marrow and adipose-derived mesenchymal stromal cells obtained from one rabbit. MSCs from both sources display the characteristic spindle-shaped morphology. Magnification ×4. B Immunophenotype of culture-expanded ASCs obtained from one representative rabbit. ASCs were positive for CD90, CD29 and CD49e and negative for CD45 and CD10, as reported [20]. The immunophenotype of BM-MSCs was superimposable. C Osteogenic and adipogenic differentiation capacity of BM-MSCs and ASC. Differentiation into osteoblasts was demonstrated by the histological detection of Alkaline phosphatase activity (a) and calcium depositions positive for Alizarin Red (b); magnification ×20. Differentiation into adipocytes was revealed by the formation of lipid droplets stained with Oil Red O (c); magnification ×20. Non differentiated cells (negative ctrl) are reported.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4495634&req=5

Fig2: Characterization of rabbit MSCs. A Morphology of bone marrow and adipose-derived mesenchymal stromal cells obtained from one rabbit. MSCs from both sources display the characteristic spindle-shaped morphology. Magnification ×4. B Immunophenotype of culture-expanded ASCs obtained from one representative rabbit. ASCs were positive for CD90, CD29 and CD49e and negative for CD45 and CD10, as reported [20]. The immunophenotype of BM-MSCs was superimposable. C Osteogenic and adipogenic differentiation capacity of BM-MSCs and ASC. Differentiation into osteoblasts was demonstrated by the histological detection of Alkaline phosphatase activity (a) and calcium depositions positive for Alizarin Red (b); magnification ×20. Differentiation into adipocytes was revealed by the formation of lipid droplets stained with Oil Red O (c); magnification ×20. Non differentiated cells (negative ctrl) are reported.
Mentions: ASCs and BM-MSCs showed the typical spindle shape morphology (Figure 2, Panel A) and they resulted positive for CD49e, CD90 and CD29, while they were negative for CD45 and CD10 (Figure 2, Panel B for in vitro-expanded ASCs) as reported by Piccinno et al. [21].Figure 2

Bottom Line: Rabbit ASCs were isolated and expanded in vitro with relative abundance, cells expressed typical surface markers (CD49e, CD90 and CD29).Improved re-epithelization, reduced inflammatory infiltration and increased collagen deposition were observed in biopsies from wounds treated with ASCs, with the best result in the autologous setting.Pre-clinical studies are however necessary to validate the best skin regeneration technique, which could be used in pediatric surgical translational research.

View Article: PubMed Central - PubMed

Affiliation: Pediatric Surgery Unit, Fondazione IRCCS Policlinico San Matteo and University of Pavia, 27100, Pavia, Italy. g.pelizzo@smatteo.pv.it.

ABSTRACT

Objective: Mesenchymal stromal cells (MSCs) expanded in vitro have been proposed as a potential therapy for congenital or acquired skin defects in pediatrics. The aim of this pre-clinical study was to investigate the effects of intradermal injections of MSC in experimental cutaneous wound repair comparing allogeneic and autologous adipose stem cells (ASCs) and autologous bone marrow-mesenchymal stromal cells (BM-MSCs).

Methods: Mesenchymal stromal cells were in vitro expanded from adipose and BM tissues of young female New Zealand rabbits. MSCs were characterized for plastic adhesion, surface markers, proliferation and differentiation capacity. When an adequate number of cells (ASCs 10 × 10(6) and BM-MSCs 3 × 10(6), because of their low rate of proliferation) was reached, two skin wounds were surgically induced in each animal. The first was topically treated with cell infusions, the second was used as a control. The intradermal inoculation included autologous or allogeneic ASCs or autologous BM-MSCs. For histological examination, animals were sacrificed and wounds were harvested after 11 and 21 days of treatment.

Results: Rabbit ASCs were isolated and expanded in vitro with relative abundance, cells expressed typical surface markers (CD49e, CD90 and CD29). Topically, ASC inoculation provided more rapid wound healing than BM-MSCs and controls. Improved re-epithelization, reduced inflammatory infiltration and increased collagen deposition were observed in biopsies from wounds treated with ASCs, with the best result in the autologous setting. ASCs also improved restoration of skin architecture during wound healing.

Conclusion: The use of ASCs may offer a promising solution to treat extended wounds. Pre-clinical studies are however necessary to validate the best skin regeneration technique, which could be used in pediatric surgical translational research.

No MeSH data available.


Related in: MedlinePlus