EXPERIMENTAL STUDY ON HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS-ALGINATE WOUND DRESSING_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

WANGSong ¹ , SUMeilan ² , YANGHuachao ¹ , LONGGang ¹ , TANGZhenrui ¹ ,  HUANGWen ¹

1. Department of Vascular Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, P. R. China;
2. Department of Psychiatry, Sanxia Central Hospital of Wanzhou District of Chongqing;

Corresponding?author：

HUANGWen, Email: dhuangwen@hotmail.com

Keywords：

Wound dressing; Human umbilical cord mesenchymal stem cells; Alginate gel; Three-dimensional culture; Vascular endothelial growth factor

DOI：

10.7507/1002-1892.20150247

Video：

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Objective To observe the growth characteristics of human umbilical cord mesenchymal stem cells (hUCMSCs) cultured on the alginate gel scaffolds and to explore the feasibility of hUCMSCs-alginate dressing for wound healing. Methods hUCMSCs were separated from human umbilical cords and cultured in vitro. After the 4th passage cells were co-cultured with alginate gel (experimental group), the cell growth characteristics were observed under the inverted phase contrast microscope. Vascular endothelial growth factor (VEGF) content was measured and the number of cells was counted at 0, 3, 6, and 9 days after culture; and the cell migration capacity was observed. The hUCMSCs were cultured without alginated gel as control. The model of full-thickness skin defects was established in 32 8-weekold Balb/c male mice and they were randomly divided into 4 groups (n=8): wounds were covered with hUCMSCsalginate gel compound (MSC-gel group), cell supernatants-alginate gel compound (CS-gel group), 10% FBS-alginate gel compound (FBS-gel group), and 0.01 mol/L PBS-alginate compound (PBS-gel group), respectively. Wound healing rates at 5, 10, and 15 days were observed and calculated; and the wound tissues were harvested for histological and immunohistochemical staining to assess new skin conditions at 15 days after operation. Results hUCMSCs grew well with grape-like proliferation on the alginate gel, but no cell migration was observed at 7 days after cultivation. VEGF expression and cell number in experimental group were significantly less than those in control group at 3 days（P<0.05）; then they gradually increased, and VEGF expression and cell number were significantly more than those in control group at 9 days (P<0.05). The wound healing rates of MSC-gel and CS-gel groups were significantly higher than those of FBSgel and PBS-gel groups at 5, 10, and 15 days (P<0.05). The squamous epithelium, fibroblasts, sebaceous glands, capillaries and VEGF expression of the new skin in MSC-gel and CS-gel groups were significantly more than FBS-gel and PBS-gel groups (P<0.05). But there was no significance between MSC-gel and CS-gel groups (P>0.05). Conclusion hUCMSCs can continuously express VEGF in alginate gel, which is necessary for wound healing. The hUCMSCs-alginate compound is probably a good wound dressing.

Citation： WANGSong, SUMeilan, YANGHuachao, LONGGang, TANGZhenrui, HUANGWen. EXPERIMENTAL STUDY ON HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS-ALGINATE WOUND DRESSING. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(9): 1137-1143. doi: 10.7507/1002-1892.20150247 Copy

1.	Galeano M, Altavilla D, Bitto A, et al. Recombinant human erythropoietin improves angiogenesis and wound healing in experimental burn wounds. Crit Care Med, 2006, 34(4):1139-1146.
2.	Triller C, Huljev D, Planinsek Rucigaj T. Modern wound dressings. Acta Med Croatica, 2013, 67 Suppl 1:81-87.
3.	Balakrishnan B, Mohanty M, Umashankar PR, et al. Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin. Biomaterials, 2005, 26(32):6335-6342.
4.	Augst AD, Kong HJ, Mooney DJ. Alginate hydrogels as biomaterials. Macromol Biosci, 2006, 6(8):623-633.
5.	Raof NA, Raja WK, Castracane J, et al. Bioengineering embryonic stem cell microenvironments for exploring inhibitory effects on metastatic breast cancer cells. Biomaterials, 2011, 32(17):4130-4139.
6.	Wang L, Shelton RM, Cooper PR, et al. Evaluation of sodium alginate for bone marrow cell tissue engineering. Biomaterials, 2003, 24(20):3475-3481.
7.	Kulseng B, Skj?k-Braek G, F?lling I, et al. TNF production from peripheral blood mononuclear cells in diabetic patients after stimulation with alginate and lipopolysaccharide. Scand J Immunol, 1996, 43(3):335-340.
8.	Kuijpers AJ, van Wachem PB, van Luyn MJ, et al. In vivo and in vitro release of lysozyme from cross-linked gelatin hydrogels: a model system for the delivery of antibacterial proteins from prosthetic heart valves. J Control Releas, 2000, 67(2-3):323-336.
9.	Bouhadir KH, Alsberg E, Mooney DJ. Hydrogels for combination delivery of antineoplastic agents. Biomaterials, 2001, 22(19):2625-2633.
10.	Bouhadir KH, Lee KY, Alsberg E, et al. Degradation of partially oxidized alginate and its potential application for tissue engineering. Biotechnol Prog, 2001, 17(5):945-950.
11.	McGuckin CP, Forraz N, Baradez MO, et al. Production of stem cells with embryonic characteristics from human umbilical cord blood. Cell proliferation, 2005, 38(4):245-255.
12.	Sabapathy V, Sundaram B, V M S, et al. Human Wharton's jelly mesenchymal stem cells plasticity augments scar-free skin wound healing with hair growth. PLoS One, 2014, 9(4):e93726.
13.	Arno AI, Amini-Nik S, Blit PH, et al. Human Wharton's jelly mesenchymal stem cells promote skin wound healing through paracrine signaling. Stem Cell Res Ther, 2014, 5(1):28.
14.	Shakespeare P. Burn wound healing and skin substitutes. Burns, 2001, 27(5):517-522.
15.	Langer R, Vacanti JP. Tissue engineering. Science, 1993, 260(5110): 920-926.
16.	Magyar JP, Nemir M, Ehler E, et al. Mass production of embryoid bodies in microbeads. Ann N Y Acad Sci, 2001, 944:135-143.
17.	Oca-Cossio J, Simpson NE, Han Z, et al. Effects of alginate encapsulation on mitochondrial activity. J Mater Sci Mater Med, 2005, 16(6):521-524.
18.	劉楊, 任力, 季培紅, 等. 海藻酸鈉的提純及海藻酸鈣多孔支架的制備. 華南理工大學學報:自然科學版, 2012, 40(7):142-147.
19.	Newman RE, Yoo D, LeRoux MA, et al. Treatment of inflammatory diseases with mesenchymal stem cells. Inflamm Allergy Drug Targets, 2009, 8(2):110-123.
20.	Yuan Y, Sin WY, Xue B, et al. Novel alginate three-dimensional static and rotating culture systems for effective ex vivo amplification of human cord blood hematopoietic stem cells and in vivo functional analysis of amplified cells in NOD/SCID mice. Transfusion, 2013, 53(9):2001-2011.
21.	Shohara R, Yamamoto A, Takikawa S, et al. Mesenchymal stromal cells of human umbilical cord Wharton's jelly accelerate wound healing by paracrine mechanisms. Cytotherapy, 2012, 14(10):1171-1181.

1. Galeano M, Altavilla D, Bitto A, et al. Recombinant human erythropoietin improves angiogenesis and wound healing in experimental burn wounds. Crit Care Med, 2006, 34(4):1139-1146.
2. Triller C, Huljev D, Planinsek Rucigaj T. Modern wound dressings. Acta Med Croatica, 2013, 67 Suppl 1:81-87.
3. Balakrishnan B, Mohanty M, Umashankar PR, et al. Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin. Biomaterials, 2005, 26(32):6335-6342.
4. Augst AD, Kong HJ, Mooney DJ. Alginate hydrogels as biomaterials. Macromol Biosci, 2006, 6(8):623-633.
5. Raof NA, Raja WK, Castracane J, et al. Bioengineering embryonic stem cell microenvironments for exploring inhibitory effects on metastatic breast cancer cells. Biomaterials, 2011, 32(17):4130-4139.
6. Wang L, Shelton RM, Cooper PR, et al. Evaluation of sodium alginate for bone marrow cell tissue engineering. Biomaterials, 2003, 24(20):3475-3481.
7. Kulseng B, Skj?k-Braek G, F?lling I, et al. TNF production from peripheral blood mononuclear cells in diabetic patients after stimulation with alginate and lipopolysaccharide. Scand J Immunol, 1996, 43(3):335-340.
8. Kuijpers AJ, van Wachem PB, van Luyn MJ, et al. In vivo and in vitro release of lysozyme from cross-linked gelatin hydrogels: a model system for the delivery of antibacterial proteins from prosthetic heart valves. J Control Releas, 2000, 67(2-3):323-336.
9. Bouhadir KH, Alsberg E, Mooney DJ. Hydrogels for combination delivery of antineoplastic agents. Biomaterials, 2001, 22(19):2625-2633.
10. Bouhadir KH, Lee KY, Alsberg E, et al. Degradation of partially oxidized alginate and its potential application for tissue engineering. Biotechnol Prog, 2001, 17(5):945-950.
11. McGuckin CP, Forraz N, Baradez MO, et al. Production of stem cells with embryonic characteristics from human umbilical cord blood. Cell proliferation, 2005, 38(4):245-255.
12. Sabapathy V, Sundaram B, V M S, et al. Human Wharton's jelly mesenchymal stem cells plasticity augments scar-free skin wound healing with hair growth. PLoS One, 2014, 9(4):e93726.
13. Arno AI, Amini-Nik S, Blit PH, et al. Human Wharton's jelly mesenchymal stem cells promote skin wound healing through paracrine signaling. Stem Cell Res Ther, 2014, 5(1):28.
14. Shakespeare P. Burn wound healing and skin substitutes. Burns, 2001, 27(5):517-522.
15. Langer R, Vacanti JP. Tissue engineering. Science, 1993, 260(5110): 920-926.
16. Magyar JP, Nemir M, Ehler E, et al. Mass production of embryoid bodies in microbeads. Ann N Y Acad Sci, 2001, 944:135-143.
17. Oca-Cossio J, Simpson NE, Han Z, et al. Effects of alginate encapsulation on mitochondrial activity. J Mater Sci Mater Med, 2005, 16(6):521-524.
18. 劉楊, 任力, 季培紅, 等. 海藻酸鈉的提純及海藻酸鈣多孔支架的制備. 華南理工大學學報:自然科學版, 2012, 40(7):142-147.
19. Newman RE, Yoo D, LeRoux MA, et al. Treatment of inflammatory diseases with mesenchymal stem cells. Inflamm Allergy Drug Targets, 2009, 8(2):110-123.
20. Yuan Y, Sin WY, Xue B, et al. Novel alginate three-dimensional static and rotating culture systems for effective ex vivo amplification of human cord blood hematopoietic stem cells and in vivo functional analysis of amplified cells in NOD/SCID mice. Transfusion, 2013, 53(9):2001-2011.
21. Shohara R, Yamamoto A, Takikawa S, et al. Mesenchymal stromal cells of human umbilical cord Wharton's jelly accelerate wound healing by paracrine mechanisms. Cytotherapy, 2012, 14(10):1171-1181.

Chinese Journal of Reparative and Reconstructive Surgery

EXPERIMENTAL STUDY ON HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS-ALGINATE WOUND DRESSING

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