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The Effect of Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Media Topically Delivered in Fibrin Glue on Chronic Wound Healing in Rats.

Mehanna RA, Nabil I, Attia N, Bary AA, Razek KA, Ahmed TA, Elsayed F - Biomed Res Int (2015)

Bottom Line: Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups.Although--later--none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1) structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson's trichrome-stained skin biopsies and (2) functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength.Thus, we conclude that topically applied BM-MSCs and their CM-via fibrin vehicle--could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure.

View Article: PubMed Central - PubMed

Affiliation: Medical Physiology Department, Faculty of Medicine, Alexandria University, Dr Fahmi Abdel Meguid Street, Mowassat Building, El Shatby, Alexandria 21561, Egypt ; Center of Excellence for Research in Regenerative Medicine and Applications (CERRMA), Faculty of Medicine, Alexandria University, Alexandria 21514, Egypt.

ABSTRACT
Bone marrow-derived mesenchymal stem cells (BM-MSCs) represent a modern approach for management of chronic skin injuries. In this work, we describe BM-MSCs application versus their conditioned media (CM) when delivered topically admixed with fibrin glue to enhance the healing of chronic excisional wounds in rats. Fifty-two adult male rats were classified into four groups after induction of large-sized full-thickness skin wound: control group (CG), fibrin only group (FG), fibrin + MSCs group (FG + SCs), and fibrin + CM group (FG + CM). Healing wounds were evaluated functionally and microscopically. Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups. Although--later--none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1) structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson's trichrome-stained skin biopsies and (2) functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength. Thus, we conclude that topically applied BM-MSCs and their CM-via fibrin vehicle--could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure.

No MeSH data available.


Related in: MedlinePlus

(a)–(d) Immunohistochemistry of granulation tissue 8 days after injury showing variable degrees of immunoreactivity in CD68+ macrophages (brown color). Each figure is a hematoxylin-counterstained representative section (Scale bar = 50 µm). (e) Macrophage count/HPF. (f)–(i) Photomicrographs of skin biopsies 35 days after injury depicting variable amounts and arrangement of collagen fibers in the dermis. Each figure is a Masson's trichrome-stained representative section (scale bar = 200 µm). (j) Histomorphometric quantification of mean collagen fraction. In (e) and (j) F-test (ANOVA) test was used to analyze data presented as mean ± SD; “∗” is significant versus CG and “#” is significant versus FG group [p < 0.001], while “°” is significant versus FG + CM [p < 0.05] (n = 3).
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fig3: (a)–(d) Immunohistochemistry of granulation tissue 8 days after injury showing variable degrees of immunoreactivity in CD68+ macrophages (brown color). Each figure is a hematoxylin-counterstained representative section (Scale bar = 50 µm). (e) Macrophage count/HPF. (f)–(i) Photomicrographs of skin biopsies 35 days after injury depicting variable amounts and arrangement of collagen fibers in the dermis. Each figure is a Masson's trichrome-stained representative section (scale bar = 200 µm). (j) Histomorphometric quantification of mean collagen fraction. In (e) and (j) F-test (ANOVA) test was used to analyze data presented as mean ± SD; “∗” is significant versus CG and “#” is significant versus FG group [p < 0.001], while “°” is significant versus FG + CM [p < 0.05] (n = 3).

Mentions: Examination of hematoxylin-counter stained sections revealed the central part of the wound with ulcerated skin surface, the dermis and subcutaneous tissue show a granulation tissue formed of small newly formed capillary-sized blood vessels lined by plump endothelial cells and proliferating fibroblasts, and the intervening stroma is edematous and shows variable numbers CD68+ macrophages admixed with other inflammatory cellular infiltrate (Figures 3(a)–3(d)). The mean number of macrophages/HPF was 39.44 ± 3.47 in both CG and FG, 69.44 ± 3.47 in FG + SCs and 59.44 ± 3.47 in FG + CM. As shown in Figure 3(e), no difference was detected between numbers of macrophages in the first two groups. However, in FG + SCs and FG + CM groups, their count was significantly higher than FG and CG groups (p < 0.01). The samples of FG + SCs group were more infiltrated by macrophages than FG + CM group (p < 0.05).


The Effect of Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Media Topically Delivered in Fibrin Glue on Chronic Wound Healing in Rats.

Mehanna RA, Nabil I, Attia N, Bary AA, Razek KA, Ahmed TA, Elsayed F - Biomed Res Int (2015)

(a)–(d) Immunohistochemistry of granulation tissue 8 days after injury showing variable degrees of immunoreactivity in CD68+ macrophages (brown color). Each figure is a hematoxylin-counterstained representative section (Scale bar = 50 µm). (e) Macrophage count/HPF. (f)–(i) Photomicrographs of skin biopsies 35 days after injury depicting variable amounts and arrangement of collagen fibers in the dermis. Each figure is a Masson's trichrome-stained representative section (scale bar = 200 µm). (j) Histomorphometric quantification of mean collagen fraction. In (e) and (j) F-test (ANOVA) test was used to analyze data presented as mean ± SD; “∗” is significant versus CG and “#” is significant versus FG group [p < 0.001], while “°” is significant versus FG + CM [p < 0.05] (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4508387&req=5

fig3: (a)–(d) Immunohistochemistry of granulation tissue 8 days after injury showing variable degrees of immunoreactivity in CD68+ macrophages (brown color). Each figure is a hematoxylin-counterstained representative section (Scale bar = 50 µm). (e) Macrophage count/HPF. (f)–(i) Photomicrographs of skin biopsies 35 days after injury depicting variable amounts and arrangement of collagen fibers in the dermis. Each figure is a Masson's trichrome-stained representative section (scale bar = 200 µm). (j) Histomorphometric quantification of mean collagen fraction. In (e) and (j) F-test (ANOVA) test was used to analyze data presented as mean ± SD; “∗” is significant versus CG and “#” is significant versus FG group [p < 0.001], while “°” is significant versus FG + CM [p < 0.05] (n = 3).
Mentions: Examination of hematoxylin-counter stained sections revealed the central part of the wound with ulcerated skin surface, the dermis and subcutaneous tissue show a granulation tissue formed of small newly formed capillary-sized blood vessels lined by plump endothelial cells and proliferating fibroblasts, and the intervening stroma is edematous and shows variable numbers CD68+ macrophages admixed with other inflammatory cellular infiltrate (Figures 3(a)–3(d)). The mean number of macrophages/HPF was 39.44 ± 3.47 in both CG and FG, 69.44 ± 3.47 in FG + SCs and 59.44 ± 3.47 in FG + CM. As shown in Figure 3(e), no difference was detected between numbers of macrophages in the first two groups. However, in FG + SCs and FG + CM groups, their count was significantly higher than FG and CG groups (p < 0.01). The samples of FG + SCs group were more infiltrated by macrophages than FG + CM group (p < 0.05).

Bottom Line: Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups.Although--later--none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1) structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson's trichrome-stained skin biopsies and (2) functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength.Thus, we conclude that topically applied BM-MSCs and their CM-via fibrin vehicle--could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure.

View Article: PubMed Central - PubMed

Affiliation: Medical Physiology Department, Faculty of Medicine, Alexandria University, Dr Fahmi Abdel Meguid Street, Mowassat Building, El Shatby, Alexandria 21561, Egypt ; Center of Excellence for Research in Regenerative Medicine and Applications (CERRMA), Faculty of Medicine, Alexandria University, Alexandria 21514, Egypt.

ABSTRACT
Bone marrow-derived mesenchymal stem cells (BM-MSCs) represent a modern approach for management of chronic skin injuries. In this work, we describe BM-MSCs application versus their conditioned media (CM) when delivered topically admixed with fibrin glue to enhance the healing of chronic excisional wounds in rats. Fifty-two adult male rats were classified into four groups after induction of large-sized full-thickness skin wound: control group (CG), fibrin only group (FG), fibrin + MSCs group (FG + SCs), and fibrin + CM group (FG + CM). Healing wounds were evaluated functionally and microscopically. Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups. Although--later--none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1) structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson's trichrome-stained skin biopsies and (2) functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength. Thus, we conclude that topically applied BM-MSCs and their CM-via fibrin vehicle--could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure.

No MeSH data available.


Related in: MedlinePlus