Meglei Anastasiia Yurevna, Nedorubova Irina Alekseevna, Basina Viktoriia Pavlovna, Chernomyrdina Viktoria Olegovna, Nedorubov Andrey Anatolevich, Kuznetsova Valeriya Sergeevna, Vasilyev Andrey Vyacheslavovich, Kutsev Sergey Ivanovich, Goldshtein Dmitry Vadimovich, Bukharova Tatiana Borisovna
Research Centre for Medical Genetics, 115478 Moscow, Russia.
Central Research Institute of Dental and Maxillofacial Surgery, 119021 Moscow, Russia.
Biomedicines. 2024 Oct 26;12(11):2461. doi: 10.3390/biomedicines12112461.
The replenishment of bone deficiency remains a challenging task in clinical practice. The use of gene-activated matrices (GAMs) impregnated with genetic constructs may be an innovative approach to solving this problem. The aim of this work is to develop collagen-based matrices with the addition of platelet-rich plasma, carrying polyplexes with the gene, to study their biocompatibility and osteogenic potential in vitro and in vivo. The cytocompatibility of the materials during incubation with adipose-derived stem cells (ADSCs) was studied using the MTT test and fluorescent microscopy. Biocompatibility was assessed during intramuscular implantation, followed by histological analysis. Osteogenic differentiation was determined by the expressions of and using real-time PCR and extracellular matrix (ECM) mineralization by alizarin red staining. The efficiency of bone regeneration was studied using micro-CT and analysis of histological sections stained according to Masson. After the incubation of ADSCs with GAS, significant increases in the expressions of the and genes by 3 ± 0.1 and 9.9 ± 0.6 times, relative to the control, as well as mineralization of the ECM, were observed. The volume of newly formed bone was 37.2 ± 6.2% after implantation of GAS, 20.9 ± 1.2%-non-activated Col/PRP, and 2.6 ± 1.5% in an empty defect. The use of Col/PRP-based matrices is an effective method for delivering of the osteoinductor gene to the site of bone tissue damage. The highest degree of healing was observed after the implantation of Col/PRP-TF/pBMP2 into the critical size defect compared to the other groups.
在临床实践中，骨缺损的修复仍然是一项具有挑战性的任务。使用浸渍有基因构建体的基因激活基质（GAMs）可能是解决这一问题的创新方法。这项工作的目的是开发添加富血小板血浆的基于胶原蛋白的基质，携带与该基因的多聚体，以研究其在体外和体内的生物相容性和成骨潜力。使用MTT试验和荧光显微镜研究了材料与脂肪来源干细胞（ADSCs）孵育期间的细胞相容性。在肌肉内植入后评估生物相容性，随后进行组织学分析。通过实时PCR检测和成骨分化，通过茜素红染色检测细胞外基质（ECM）矿化。使用微型计算机断层扫描（micro-CT）和根据Masson染色的组织学切片分析研究骨再生效率。在ADSCs与GAS孵育后，相对于对照，观察到和成骨基因的表达显著增加3±0.1倍和9.9±0.6倍，以及ECM矿化。植入GAS后新形成骨的体积为37.2±6.2%，未激活的Col/PRP为20.9±1.2%，空缺损为2.6±1.5%。使用基于Col/PRP的基质是将骨诱导基因递送至骨组织损伤部位的有效方法。与其他组相比，将Col/PRP-TF/pBMP2植入临界尺寸缺损后观察到最高程度的愈合。
