Stem Cell Research Laboratory, Cell Therapy and Biotechnology in Regenerative Medicine Department, Pequeno Príncipe Faculties and the Pelé Pequeno Príncipe Research Institute, Curitiba, Paraná, Brazil.
Chemistry Undergraduate Program, School of Education and Humanities, Pontifícia Universidade Católica Do Paraná, Curitiba, Paraná, Brazil.
Int J Nanomedicine. 2021 Feb 5;16:833-850. doi: 10.2147/IJN.S289868. eCollection 2021.
In deep burns, wound contraction and hypertrophic scar formation can generate functional derangement and debilitation of the affected part. In order to improve the quality of healing in deep second-degree burns, we developed a new treatment in a preclinical model using nanostructured membranes seeded with mesenchymal stem cells (MSCs).
Membranes were obtained by reconstitution of bacterial cellulose (reconstituted membrane [RM]) and produced by a dry-cast process, then RM was incorporated with 10% tamarind xyloglucan plus gellan gum 1:1 and 10% lysozyme (RMGT-LZ) and with 10% gellan gum and 10% lysozyme (RMG-LZ). Membrane hydrophobic/hydrophilic characteristics were investigated by static/dynamic contact-angle measurements. They were cultivated with MSCs, and cell adhesion, proliferation, and migration capacity was analyzed with MTT assays. Morphological and topographic characteristics were analyzed by scanning electron microscopy. MSC patterns in flow cytometry and differentiation into adipocytes and osteocytes were checked. In vivo assays used RMG-LZ and RMGT-LZ (with and without MSCs) in rats submitted to burn protocol, and histological sections and collagen deposits were analyzed and immunocytochemistry assay performed.
In vitro results demonstrated carboxyl and amine groups made the membranes moderately hydrophobic and xyloglucan inclusion decreased wettability, favoring MSC adhesion, proliferation, and differentiation. In vivo, we obtained 40% and 60% reduction in acute/chronic inflammatory infiltrates, 96% decrease in injury area, increased vascular proliferation and collagen deposition, and complete epithelialization after 30 days. MSCs were detected in burned tissue, confirming they had homed and proliferated in vivo.
Nanostructured cellulose-gellan-xyloglucan-lysozyme dressings, especially when seeded with MSCs, improved deep second-degree burn regeneration.
在深度烧伤中,伤口收缩和增生性瘢痕形成会导致受影响部位的功能障碍和虚弱。为了改善深度二度烧伤的愈合质量,我们在临床前模型中开发了一种使用间充质干细胞(MSCs)种植的纳米结构膜的新治疗方法。
通过再构成细菌纤维素(重组膜[RM])获得膜,并通过干法铸造工艺生产,然后将 RM 与 10%罗望子木葡聚糖加结冷胶 1:1 和 10%溶菌酶(RMGT-LZ)以及 10%结冷胶和 10%溶菌酶(RMG-LZ)结合。通过静态/动态接触角测量研究了膜的疏水性/亲水性特征。用 MSCs 培养它们,并通过 MTT 测定分析细胞黏附、增殖和迁移能力。通过扫描电子显微镜分析形态和形貌特征。通过流式细胞术检查 MSC 模式和向脂肪细胞和成骨细胞的分化。体内试验使用 RMG-LZ 和 RMGT-LZ(有/无 MSCs)在接受烧伤方案的大鼠中进行,分析组织学切片和胶原蛋白沉积,并进行免疫细胞化学测定。
体外结果表明,羧基和氨基基团使膜具有中等疏水性,木葡聚糖的包含降低了润湿性,有利于 MSC 的黏附、增殖和分化。在体内,我们获得了 40%和 60%的急性/慢性炎症浸润减少,损伤面积减少 96%,血管增殖和胶原蛋白沉积增加,30 天后完全上皮化。在烧伤组织中检测到 MSCs,证实它们已归巢并在体内增殖。
纳米结构纤维素-结冷胶-木葡聚糖-溶菌酶敷料,特别是当种植 MSCs 时,改善了深度二度烧伤的再生。