Tissue-engineered skin for skin wound repair: construction by human acellular dermal matrix combined with bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.0889

Abstract

Abstract:

BACKGROUND: A variety of seed cells and scaffolds can be used for constructing tissue-engineered skin. Exploring a proper construction method is a hot topic of research.

OBJECTIVE: To explore the feasibility of tissue-engineered skin to promote healing of skin wounds in rats.

METHODS: Sprague-Dawley rats of 6-8 weeks old were taken to isolate bone marrow mesenchymal stem cells (BMSCs) using the whole bone marrow adherent method. Then, BMSCs were cultured and purified in vitro. Human foreskin samples were digested using enzyme digestion method to make human acellular dermal matrix (ADM). Tissue-engineered skin was thereafter constructed using BMSCs as seed cells and ADM as the scaffold, to cover the skin defect wound in the rats. Skin wound covered with ADM and nothing was used as ADM and blank control groups, respectively. Wound healing time was recorded and wound closure index was calculated. Evaluation of wound healing was conducted through hematoxylin-eosin, CK19 and BrdU immunohistochemical staining of wound samples at 2 weeks after transplantation.

RESULTS AND CONCLUSION: The wound closure indexes in the three groups were ranked as follows: tissue-engineered skin group > ADM group > blank control group, and there were significant differences between groups (P < 0.05). The wound healing time was shortest in the tissue-engineered skin group, shorter in the ADM group and longest in the blank control group, and there were also significant differences between groups (P < 0.05). Cells positive for BrdU were detected in the tissue-engineered skin group, and the expression level of CK19 was significantly higher in the tissue-engineered skin group as compared with the other two groups. In conclusion, the tissue-engineered skin constructed by BMSCs and ADM achieves good outcomes in the repair of full-thickness skin wound.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Skin Transplantation, Wound Healing, Tissue Engineering

CLC Number:

R318

Chi Kai, Wu Lei, Chen Long-jin, Li Yong-lin. Tissue-engineered skin for skin wound repair: construction by human acellular dermal matrix combined with bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(26): 4179-4183.

Figures/Tables 7

References

[1] 陈璧,姜笃银,贾赤宇,等.复合皮移植的实验研究与临床应用[J].中华烧伤杂志,2004,20(6):347-350.

[2] Duffy MM, Ritter T, Ceredig R, et al. Mesenchymal stem cell effects on T-cell effector pathways. Stem Cell Res Ther. 2011; 2(4):34.

[3] Delirezh N, Shojaeefar E, Parvin P, et al. Comparison the effects of two monocyte isolation methods, plastic adherence and magnetic activated cell sorting methods, on phagocytic activity of generated dendritic cells. Cell J. 2013;15(3): 218-223.

[4] 李晓峰,赵劲民,苏伟,等.大鼠骨髓间充质干细胞的培养与鉴定[J].中国组织工程研究与临床康复,2011,15(10):1721-1725.

[5] 刘坡,祁少海,舒斌,等.异体脱细胞真皮基质作为组织工程皮肤真皮支架的可行性[J].中国组织工程研究, 2012,16(21): 3864-3868.

[6] Alhadlaq A, Mao JJ. Mesenchymal stem cells: isolation and therapeutics. Stem Cells Dev. 2004;13(4):436-448.

[7] Lee RH, Kim B, Choi I, et al. Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue. Cell Physiol Biochem. 2004;14 (4-6):311-324.

[8] Rasini V, Dominici M, Kluba T, et al. Mesenchymal stromal/ stem cells markers in the human bone marrow. Cytotherapy. 2013;15(3):292-306.

[9] Lu X, Alshemali S, de Wynter EA, et al. Mesenchymal stem cells from CD34(-) human umbilical cord blood. Transfus Med. 2010;20(3):178-184.

[10] 韩跃东,刘玲.组织工程皮肤的临床应用[J].国际皮肤性病学杂志志,2007,33(6):340-342.

[11] Mirastschijski U, Bugdahl R, Rollman O, et al. Epithelial regeneration from bioengineered skin explants in culture. Br J Dermatol. 2006;154(1):42-49.

[12] 迟作华,张洹,陆琰,等.人骨髓间充质干细胞分离方法的比较[J].中国临床康复,2006,10(1):20-22.

[13] Pereira RF, Halford KW, O'Hara MD, et al. Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. Proc Natl Acad Sci U S A. 1995;92(11):4857-4861.

[14] Zhang B, Wang F, Deng L, et al. Isolating and culturing rat marrow mesenchymal stem cells and studying their phenotypical and functional properties. Sichuan Da Xue Xue Bao Yi Xue Ban. 2003;34(4):738-741.

[15] Boiret N, Rapatel C, Veyrat-Masson R, et al. Characterization of nonexpanded mesenchymal progenitor cells from normal adult human bone marrow. Exp Hematol. 2005;33(2): 219-225.

[16] Jones EA, English A, Kinsey SE, et al. Optimization of a flow cytometry-based protocol for detection and phenotypic characterization of multipotent mesenchymal stromal cells from human bone marrow. Cytometry B Clin Cytom. 2006; 70(6):391-399.

[17] Gao K, Lu Y, Li S, et al. Isolation, culturing and growth characteristics of mesenchymal stem cells from bone marrow of Rhesus monkey, Macaca mulatta. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2007;24(6):1343-1347, 1351.

[18] Nadri S, Soleimani M, Hosseni RH, et al. An efficient method for isolation of murine bone marrow mesenchymal stem cells. Int J Dev Biol. 2007;51(8):723-729.