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Microencapsulated equine mesenchymal stromal cells promote cutaneous wound healing in vitro.

Bussche L, Harman RM, Syracuse BA, Plante EL, Lu YC, Curtis TM, Ma M, Van de Walle GR - Stem Cell Res Ther (2015)

Bottom Line: Mesenchymal stromal cells (MSCs) have been reported to provide paracrine signals that promote wound healing, but (i) how they exert their effects on target cells is unclear and (ii) a suitable delivery system to supply these MSC-derived secreted factors in a controlled and safe way is unavailable.In addition, we found that equine MSCs packaged in core-shell hydrogel microcapsules had similar effects on equine dermal fibroblast migration and gene expression, indicating that microencapsulation of MSCs does not interfere with the release of bioactive factors.Our results demonstrate that the use of CM from MSCs might be a promising new therapy for impaired cutaneous wounds and that encapsulation may be a suitable way to effectively deliver CM to wounded cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, 235 Hungerford Hill Road, Ithaca, NY, 14850, USA. lb548@cornell.edu.

ABSTRACT

Introduction: The prevalence of impaired cutaneous wound healing is high and treatment is difficult and often ineffective, leading to negative social and economic impacts for our society. Innovative treatments to improve cutaneous wound healing by promoting complete tissue regeneration are therefore urgently needed. Mesenchymal stromal cells (MSCs) have been reported to provide paracrine signals that promote wound healing, but (i) how they exert their effects on target cells is unclear and (ii) a suitable delivery system to supply these MSC-derived secreted factors in a controlled and safe way is unavailable. The present study was designed to provide answers to these questions by using the horse as a translational model. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC-derived conditioned medium (CM), containing all factors secreted by MSCs, on equine dermal fibroblasts, a cell type critical for successful wound healing, and (ii) explore the potential of microencapsulated equine MSCs to deliver CM to wounded cells in vitro.

Methods: MSCs were isolated from the peripheral blood of healthy horses. Equine dermal fibroblasts from the NBL-6 (horse dermal fibroblast cell) line were wounded in vitro, and cell migration and expression levels of genes involved in wound healing were evaluated after treatment with MSC-CM or NBL-6-CM. These assays were repeated by using the CM collected from MSCs encapsulated in core-shell hydrogel microcapsules.

Results: Our salient findings were that equine MSC-derived CM stimulated the migration of equine dermal fibroblasts and increased their expression level of genes that positively contribute to wound healing. In addition, we found that equine MSCs packaged in core-shell hydrogel microcapsules had similar effects on equine dermal fibroblast migration and gene expression, indicating that microencapsulation of MSCs does not interfere with the release of bioactive factors.

Conclusions: Our results demonstrate that the use of CM from MSCs might be a promising new therapy for impaired cutaneous wounds and that encapsulation may be a suitable way to effectively deliver CM to wounded cells in vivo.

No MeSH data available.


Related in: MedlinePlus

Mesenchymal stromal cell (MSC) conditioned medium (CM) promotes migration of dermal fibroblasts in scratch assays (n = 3). (A) Representative phase-contrast images of wounded NBL-6 cells cultured with MSC-derived CM (lower panels) as compared with control CM (upper panels) over a 48-hour period, and migration distances of NBL-6 cells are expressed as micrometer per hour in 48 hours. **P <0.01. (B) Migration of NBL-6 cells cultured with MSC-derived CM as compared with CM from mitomycin C-treated MSCs (2,000 ng/mL). (C) Viability (dashed line) and proliferation (solid line) of NBL-6 cells cultured in the presence of various concentrations of mitomycin C. NBL-6, horse dermal fibroblast cell; OD, optical density.
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Fig1: Mesenchymal stromal cell (MSC) conditioned medium (CM) promotes migration of dermal fibroblasts in scratch assays (n = 3). (A) Representative phase-contrast images of wounded NBL-6 cells cultured with MSC-derived CM (lower panels) as compared with control CM (upper panels) over a 48-hour period, and migration distances of NBL-6 cells are expressed as micrometer per hour in 48 hours. **P <0.01. (B) Migration of NBL-6 cells cultured with MSC-derived CM as compared with CM from mitomycin C-treated MSCs (2,000 ng/mL). (C) Viability (dashed line) and proliferation (solid line) of NBL-6 cells cultured in the presence of various concentrations of mitomycin C. NBL-6, horse dermal fibroblast cell; OD, optical density.

Mentions: Because dermal fibroblasts are critically involved in wound healing [29], we decided to study the effects of MSC-derived CM on the migration of these cells by using in vitro scratch assays [30]. We found that equine dermal fibroblast cells (NBL-6) migrated significantly faster when cultured in CM obtained from equine MSCs compared with control CM, which was collected from NBL-6 (Figure 1A).Figure 1


Microencapsulated equine mesenchymal stromal cells promote cutaneous wound healing in vitro.

Bussche L, Harman RM, Syracuse BA, Plante EL, Lu YC, Curtis TM, Ma M, Van de Walle GR - Stem Cell Res Ther (2015)

Mesenchymal stromal cell (MSC) conditioned medium (CM) promotes migration of dermal fibroblasts in scratch assays (n = 3). (A) Representative phase-contrast images of wounded NBL-6 cells cultured with MSC-derived CM (lower panels) as compared with control CM (upper panels) over a 48-hour period, and migration distances of NBL-6 cells are expressed as micrometer per hour in 48 hours. **P <0.01. (B) Migration of NBL-6 cells cultured with MSC-derived CM as compared with CM from mitomycin C-treated MSCs (2,000 ng/mL). (C) Viability (dashed line) and proliferation (solid line) of NBL-6 cells cultured in the presence of various concentrations of mitomycin C. NBL-6, horse dermal fibroblast cell; OD, optical density.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4413990&req=5

Fig1: Mesenchymal stromal cell (MSC) conditioned medium (CM) promotes migration of dermal fibroblasts in scratch assays (n = 3). (A) Representative phase-contrast images of wounded NBL-6 cells cultured with MSC-derived CM (lower panels) as compared with control CM (upper panels) over a 48-hour period, and migration distances of NBL-6 cells are expressed as micrometer per hour in 48 hours. **P <0.01. (B) Migration of NBL-6 cells cultured with MSC-derived CM as compared with CM from mitomycin C-treated MSCs (2,000 ng/mL). (C) Viability (dashed line) and proliferation (solid line) of NBL-6 cells cultured in the presence of various concentrations of mitomycin C. NBL-6, horse dermal fibroblast cell; OD, optical density.
Mentions: Because dermal fibroblasts are critically involved in wound healing [29], we decided to study the effects of MSC-derived CM on the migration of these cells by using in vitro scratch assays [30]. We found that equine dermal fibroblast cells (NBL-6) migrated significantly faster when cultured in CM obtained from equine MSCs compared with control CM, which was collected from NBL-6 (Figure 1A).Figure 1

Bottom Line: Mesenchymal stromal cells (MSCs) have been reported to provide paracrine signals that promote wound healing, but (i) how they exert their effects on target cells is unclear and (ii) a suitable delivery system to supply these MSC-derived secreted factors in a controlled and safe way is unavailable.In addition, we found that equine MSCs packaged in core-shell hydrogel microcapsules had similar effects on equine dermal fibroblast migration and gene expression, indicating that microencapsulation of MSCs does not interfere with the release of bioactive factors.Our results demonstrate that the use of CM from MSCs might be a promising new therapy for impaired cutaneous wounds and that encapsulation may be a suitable way to effectively deliver CM to wounded cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, 235 Hungerford Hill Road, Ithaca, NY, 14850, USA. lb548@cornell.edu.

ABSTRACT

Introduction: The prevalence of impaired cutaneous wound healing is high and treatment is difficult and often ineffective, leading to negative social and economic impacts for our society. Innovative treatments to improve cutaneous wound healing by promoting complete tissue regeneration are therefore urgently needed. Mesenchymal stromal cells (MSCs) have been reported to provide paracrine signals that promote wound healing, but (i) how they exert their effects on target cells is unclear and (ii) a suitable delivery system to supply these MSC-derived secreted factors in a controlled and safe way is unavailable. The present study was designed to provide answers to these questions by using the horse as a translational model. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC-derived conditioned medium (CM), containing all factors secreted by MSCs, on equine dermal fibroblasts, a cell type critical for successful wound healing, and (ii) explore the potential of microencapsulated equine MSCs to deliver CM to wounded cells in vitro.

Methods: MSCs were isolated from the peripheral blood of healthy horses. Equine dermal fibroblasts from the NBL-6 (horse dermal fibroblast cell) line were wounded in vitro, and cell migration and expression levels of genes involved in wound healing were evaluated after treatment with MSC-CM or NBL-6-CM. These assays were repeated by using the CM collected from MSCs encapsulated in core-shell hydrogel microcapsules.

Results: Our salient findings were that equine MSC-derived CM stimulated the migration of equine dermal fibroblasts and increased their expression level of genes that positively contribute to wound healing. In addition, we found that equine MSCs packaged in core-shell hydrogel microcapsules had similar effects on equine dermal fibroblast migration and gene expression, indicating that microencapsulation of MSCs does not interfere with the release of bioactive factors.

Conclusions: Our results demonstrate that the use of CM from MSCs might be a promising new therapy for impaired cutaneous wounds and that encapsulation may be a suitable way to effectively deliver CM to wounded cells in vivo.

No MeSH data available.


Related in: MedlinePlus