Manso Gabriel Martins da Costa, Elias-Oliveira Jefferson, Guimarães Jhefferson Barbosa, Pereira Ítalo Sousa, Rodrigues Vanessa Fernandes, Burger Beatriz, Fantacini Daianne Maciely Carvalho, de Souza Lucas Eduardo Botelho, Rodrigues Hosana Gomes, Bonato Vânia Luiza Deperon, Silva João Santana, Ramos Simone Gusmão, Tostes Rita Cassia, Manfiolli Adriana Oliveira, Caliari-Oliveira Carolina, Carlos Daniela
Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil.
Laboratory of Nutrients and Tissue Repair, School of Applied Sciences, University of Campinas, Limeira, Brazil.
Regen Ther. 2023 Jan 9;22:79-89. doi: 10.1016/j.reth.2022.12.006. eCollection 2023 Mar.
Diabetes mellitus (DM) is a chronic disease and a major cause of mortality and morbidity worldwide. The hyperglycemia caused by DM induces micro and macrovascular complications that lead, among other consequences, to chronic wounds and amputations. Cell therapy and tissue engineering constitute recent therapeutic alternatives to improve wound healing in diabetic patients. The current study aimed to analyze the effectiveness of biocuratives containing human mesenchymal stem cells (MSCs) associated with a hydrogel matrix in the wound healing process and related inflammatory cell profile in diabetic mice.
Biocuratives containing MSCs were constructed by 3D bioprinting, and applied to skin wounds on the back of streptozotocin (STZ)-induced type 1 diabetic (T1D) mice. The healing process, after the application of biocuratives with or without MSCs was histologically analyzed. In parallel, genes related to growth factors, mast cells (MC), M1 and M2 macrophage profiles were evaluated by RT-PCR. Macrophages were characterized by flow cytometry, and MC by toluidine blue staining and flow cytometry.
Mice with T1D exhibited fewer skin MC and delayed wound healing when compared to the non-diabetic group. Treatment with the biocuratives containing MSCs accelerated wound healing and improved skin collagen deposition in diabetic mice. Increased TGF-β gene expression and M2 macrophage-related markers were also detected in skin of diabetic mice that received MSCs-containing biocuratives. Finally, MSCs upregulated IL-33 gene expression and augmented the number of MC in the skin of diabetic mice.
These results reveal the therapeutic potential of biocuratives containing MSCs in the healing of skin wounds in diabetic mice, providing a scientific base for future treatments in diabetic patients.
糖尿病（DM）是一种慢性疾病，也是全球范围内死亡率和发病率的主要原因。糖尿病引起的高血糖会诱发微血管和大血管并发症，进而导致慢性伤口和截肢等后果。细胞疗法和组织工程是近年来用于改善糖尿病患者伤口愈合的治疗选择。本研究旨在分析含人间充质干细胞（MSCs）与水凝胶基质的生物治疗剂在糖尿病小鼠伤口愈合过程及相关炎症细胞谱中的有效性。
通过3D生物打印构建含MSCs的生物治疗剂，并将其应用于链脲佐菌素（STZ）诱导的1型糖尿病（T1D）小鼠背部的皮肤伤口。对应用含或不含MSCs的生物治疗剂后的愈合过程进行组织学分析。同时，通过逆转录聚合酶链反应（RT-PCR）评估与生长因子、肥大细胞（MC）、M1和M2巨噬细胞谱相关的基因。通过流式细胞术对巨噬细胞进行表征，通过甲苯胺蓝染色和流式细胞术对MC进行表征。
与非糖尿病组相比，T1D小鼠的皮肤MC较少，伤口愈合延迟。用含MSCs的生物治疗剂治疗可加速糖尿病小鼠的伤口愈合并改善皮肤胶原蛋白沉积。在接受含MSCs生物治疗剂的糖尿病小鼠皮肤中还检测到转化生长因子-β（TGF-β）基因表达增加以及与M2巨噬细胞相关的标志物。最后，MSCs上调白细胞介素-33（IL-33）基因表达并增加糖尿病小鼠皮肤中MC的数量。
这些结果揭示了含MSCs的生物治疗剂在糖尿病小鼠皮肤伤口愈合中的治疗潜力，为未来糖尿病患者的治疗提供了科学依据。
