Chemokine receptor 7-bone marrow mesenchymal stem cells combined with porcine small intestinal submucosa promote skin repair in rats

doi:10.12307/2026.058

Abstract

Abstract: BACKGROUND: The clinical outcomes of autologous and allogeneic skin transplants, which are commonly used for repairing skin lesions, are often suboptimal. In recent years, advancements in tissue engineering have provided new hope for skin repair. Nevertheless, the regeneration of blood vessels within skin tissue engineering remains a significant challenge. Porcine small intestinal submucosa and bone marrow-derived mesenchymal stem cells are widely utilized natural extracellular matrix biomaterials and seed cells in current tissue engineering research. Chemokine receptor 7 is a cytokine that can promote angiogenesis.
OBJECTIVE: To observe the repair effect and angiogenesis ability of chemokine receptor 7-bone marrow mesenchymal stem cells-porcine small intestinal submucosa membrane on rat back skin damage.
METHODS: (1) The adenovirus vector overexpressing chemokine receptor 7 was used to transfect bone marrow mesenchymal stem cells. The transfection efficiency was evaluated using western blot assay and RT-qPCR. (2) The bone marrow mesenchymal stem cells overexpressing chemokine receptor 7 were co-cultured with porcine small intestinal submucosa. Cytocompatibility was assessed through scanning electron microscopy and live/dead cell staining. (3) 12 SD rats were utilized to establish a skin defect animal experimental model, and chemokine receptor 7-bone marrow mesenchymal stem cells-porcine small intestinal submucosa (experimental group) and porcine small intestinal submucosa (control group) were placed in the rat skin defect, and the wound healing was observed 1, 3, 7, and 14 days after modeling. The expression of vascular endothelial growth factor protein in the wound healing tissue was detected by western blot assay at 7 and 14 days after modeling, and the expression of CD31 and proliferating cell nuclear antigen protein in the wound tissue was detected by immunohistochemical staining.
RESULTS AND CONCLUSION: (1) Bone marrow mesenchymal stem cells overexpressing chemokine receptor 7 were successfully constructed by adenovirus-mediated transfection. The protein and mRNA expressions of chemokine receptor 7 in the chemokine receptor 7 transfected group were significantly upregulated compared with those in the control group and the empty vector group (P < 0.001). (2) Scanning electron microscopy observation and live-dead cell staining results showed that bone marrow mesenchymal stem cells overexpressing chemokine receptor 7 grew well on the surface of the submucosal membrane of the porcine small intestine, and the two had good compatibility. (3) Compared with the control group, the wound area of the experimental group was significantly reduced (P < 0.05). The protein expression levels of vascular endothelial growth factor, CD31, and proliferating cell nuclear antigen in the wound healing tissue of the experimental group were higher than those in the control group (P < 0.05), indicating that the ability of chemokine receptor 7-bone marrow mesenchymal stem cells-porcine small intestinal submucosal membrane to promote wound angiogenesis was strong.

Key words: chemokine receptor 7, bone marrow derived mesenchymal stem cell, adenovirus, porcine small intestinal submucosa, skin repair, angiogenesis

CLC Number:

Fan Meirong, Li Guangqi, Song Xumei, Yan Xin, Sui Ruizhi. Chemokine receptor 7-bone marrow mesenchymal stem cells combined with porcine small intestinal submucosa promote skin repair in rats[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(13): 3242-3249.

Figures/Tables 6

References

[1] HOLL J, KOWALEWSKI C, ZIMEK Z, et al. Chronic Diabetic Wounds and Their Treatment with Skin Substitutes. Cells. 2021;10(3):655.
[2] SHI CR, FERREIRA AL, KAUR M, et al. Cutaneous Chronic Graft-Versus-Host Disease: Clinical Manifestations, Diagnosis, Management, and Supportive Care. Transplant Cell Ther. 2024;30(9S):S513-S533.
[3] OZHATHIL DK, TAY MW, WOLF SE, et al. A Narrative Review of the History of Skin Grafting in Burn Care. Medicina (Kaunas). 2021;57(4):380.
[4] KHAN AA, KHAN IM, NGUYEN PP, et al. Skin Graft Techniques. Clin Podiatr Med Surg. 2020;37(4):821-835.
[5] COSTELLO L, DICOLANDREA T, TASSEFF R, et al. Tissue engineering strategies to bioengineer the ageing skin phenotype in vitro. Aging Cell. 2022;21(2):e13550.
[6] CHEN J, FAN Y, DONG G, et al. Designing biomimetic scaffolds for skin tissue engineering. Biomater Sci. 2023;11(9):3051-3076.
[7] ZHAO Y, PENG H, SUN L, et al. The application of small intestinal submucosa in tissue regeneration. Mater Today Bio. 2024;26:101032.
[8] FUJII M, TANAKA R. Porcine Small Intestinal Submucosa Alters the Biochemical Properties of Wound Healing: A Narrative Review. Biomedicines. 2022;10(9):2213.
[9] JELODARI S, SADRODDINY E. Decellularization of Small Intestinal Submucosa. Adv Exp Med Biol. 2021;1345:71-84.
[10] SUHANDI C, MOHAMMED AFA, WILAR G, et al. Effectiveness of Mesenchymal Stem Cell Secretome on Wound Healing: A Systematic Review and Meta-analysis. Tissue Eng Regen Med. 2023;20(7):1053-1062.
[11] SUKMANA BI, MARGIANA R, ALMAJIDI YQ, et al. Supporting wound healing by mesenchymal stem cells (MSCs) therapy in combination with scaffold, hydrogel, and matrix; State of the art. Pathol Res Pract. 2023;248:154575.
[12] POLTAVETS V, FAULKNER JW, DHATRAK D, et al. CXCR4-CCR7 Heterodimerization Is a Driver of Breast Cancer Progression. Life (Basel). 2021;11(10):1049.
[13] VAHEDI L, SHEIDAEI S, GHASEMI M, et al. Cytoplasmic CCR7 (CCR7c) Immunoexpression Is Associated with Tumor Invasion in Gastric Cancer. Int J Hematol Oncol Stem Cell Res. 2023;17(4):267-274.
[14] RIZEQ B, MALKI MI. The Role of CCL21/CCR7 Chemokine Axis in Breast Cancer Progression. Cancers (Basel). 2020;12(4):1036.
[15] HE J, ZHANG X, XIA X, et al. Organoid technology for tissue engineering. J Mol Cell Biol. 2020;12(8):569-579.
[16] SPEED OE, BAREISS A, PATEL VA, et al. Otologic use of porcine small intestinal submucosal graft (biodesign): A MAUDE database review. Am J Otolaryngol. 2023;44(5):103961.
[17] ZANG C, XIAN H, ZHANG H, et al. Clinical outcomes of a novel porcine small intestinal submucosa patch for full-thickness hand skin defects: a retrospective investigation. J Orthop Surg Res. 2023;18(1):50.
[18] JEFFERY S. Clinical benefits of small intestinal submucosa extracellular matrix and review of the evidence. J Wound Care. 2023;32(Sup2):S11-S19.
[19] GUO X, XIA B, LU XB, et al. Grafting of mesenchymal stem cell-seeded small intestinal submucosa to repair the deep partial-thickness burns. Connect Tissue Res. 2016;57(5):388-397.
[20] LEE C, SHIM S, JANG H, et al. Human umbilical cord blood-derived mesenchymal stromal cells and small intestinal submucosa hydrogel composite promotes combined radiation-wound healing of mice. Cytotherapy. 2017;19(9):1048-1059.
[21] FAN MR, GONG M, DA LC, et al. Tissue engineered esophagus scaffold constructed with porcine small intestinal submucosa and synthetic polymers. Biomed Mater. 2014;9(1):015012.
[22] ROSHANGAR L, SOLEIMANI RAD J, KHEIRJOU R, et al. Skin Burns: Review of Molecular Mechanisms and Therapeutic Approaches. Wounds. 2019; 31(12):308-315.
[23] PEÑA OA, MARTIN P. Cellular and molecular mechanisms of skin wound healing. Nat Rev Mol Cell Biol. 2024;25(8):599-616.
[24] LIU L, ZHENG CX, ZHAO N, et al. Mesenchymal Stem Cell Aggregation-Released Extracellular Vesicles Induce CD31+ EMCN+ Vessels in Skin Regeneration and Improve Diabetic Wound Healing. Adv Healthc Mater. 2023;12(20):e2300019.
[25] WEI Q, WANG Y, MA K, et al. Extracellular Vesicles from Human Umbilical Cord Mesenchymal Stem Cells Facilitate Diabetic Wound Healing Through MiR-17-5p-mediated Enhancement of Angiogenesis. Stem Cell Rev Rep. 2022;18(3):1025-1040.
[26] HONG W, YANG B, HE Q, et al. New Insights of CCR7 Signaling in Dendritic Cell Migration and Inflammatory Diseases. Front Pharmacol. 2022;13: 841687.
[27] GERALDO LH, GARCIA C, XU Y, et al. CCL21-CCR7 signaling promotes microglia/macrophage recruitment and chemotherapy resistance in glioblastoma. Cell Mol Life Sci. 2023;80(7):179.
[28] CAI QY, LIANG GY, ZHENG YF, et al. CCR7 enhances the angiogenic capacity of esophageal squamous carcinoma cells in vitro via activation of the NF-κB/VEGF signaling pathway. Am J Transl Res. 2017;9(7):3282-3292.
[29] YUAN LH, CHEN XL, DI Y, et al. CCR7/p-ERK1/2/VEGF signaling promotes retinal neovascularization in a mouse model of oxygen-induced retinopathy. Int J Ophthalmol. 2017;10(6):862-869.
[30] NAITO H, IBA T, TAKAKURA N. Mechanisms of new blood-vessel formation and proliferative heterogeneity of endothelial cells. Int Immunol. 2020; 32(5):295-305.
[31] AHMAD A, NAWAZ MI. Molecular mechanism of VEGF and its role in pathological angiogenesis. J Cell Biochem. 2022;123(12):1938-1965.
[32] QUEISSER A, SERONT E, BOON LM, et al. Genetic Basis and Therapies for Vascular Anomalies. Circ Res. 2021;129(1):155-173.
[33] DI BENEDETTO P, RUSCITTI P, BERARDICURTI O, et al. Blocking Jak/STAT signalling using tofacitinib inhibits angiogenesis in experimental arthritis. Arthritis Res Ther. 2021;23(1):213.
[34] VIMALRAJ S. A concise review of VEGF, PDGF, FGF, Notch, angiopoietin, and HGF signalling in tumor angiogenesis with a focus on alternative approaches and future directions. Int J Biol Macromol. 2022;221:1428-1438.
[35] FU T, SULLIVAN DP, GONZALEZ AM, et al. Mechanotransduction via endothelial adhesion molecule CD31 initiates transmigration and reveals a role for VEGFR2 in diapedesis. Immunity. 2023;56(10):2311-2324.e6.
[36] ZENG Z, CHEN H, CAI J, et al. IL-10 regulates the malignancy of hemangioma-derived endothelial cells via regulation of PCNA. Arch Biochem Biophys. 2020;688:108404.