I Gde Rurus Suryawan, Andrianto, Arisya Agita, Anudya Kartika Ratri, Ricardo Adrian Nugraha
{"title":"Emerging therapeutic benefit of platelet-rich fibrin as novel platelet concentrates in tissue engineering.","authors":"I Gde Rurus Suryawan, Andrianto, Arisya Agita, Anudya Kartika Ratri, Ricardo Adrian Nugraha","doi":"10.21542/gcsp.2024.46","DOIUrl":null,"url":null,"abstract":"<p><p><b>Background:</b> Treating irreversible cardiomyocyte loss following myocardial infarction presents several therapeutic challenges. While cell therapy shows promise as a regenerative treatment for infarcted cardiac tissue, different cell sources vary in their therapeutic potential. Adipose-derived stem cells (ADSCs) have emerged as an attractive option due to their accessibility, but their limited differentiation capacity remains a significant constraint. Recent evidence suggests that injectable platelet-rich fibrin may enhance this process by stimulating the differentiation of ADSCs into cardiomyocyte-like cells. <b>Objective:</b> Analyse the benefit of injectable platelet-rich fibrin to accelerate the differentiation of adipose-derived mesenchymal stem cells into cardiomyocyte-like cells. <b>Methods:</b> This study is a true experimental randomized pos <i>t</i>-test design study. Adipose-derived mesenchymal stem cells were isolated from adipose tissue and expanded in culture through four passages. The characteristics of adipose-derived mesenchymal stem cells were measured by the expression of CD 34-, CD 45-, and CD 105+ using flowcytometry. The samples were divided into 3 groups, i.e., negative control (<i>α</i>-MEM), positive control (differentiation medium) and treatment group (platelet-rich fibrin). The assessment of GATA-4 marker expression was conducted using flowcytometry on the fifth day and troponin was conducted using immunocytochemistry on the tenth day to determine the differentiation to cardiomyocyte. Statistical analysis was performed using Student's t-tests and one-way ANOVA for data that demonstrated normal distribution as verified by the Shapiro-Wilk test. <b>Results:</b> Flowcytometry on GATA-4 expression revealed significant difference on addition of platelet-rich fibrin compared with negative and positive controls (68.20 ± 6.82 <i>vs</i> 58.15 ± 1.23; <i>p</i> < 0.05; 68.20 ± 6.82 <i>vs</i> 52.96 ± 2.02; <i>p</i> < 0.05). This was supported by the results of immunocytochemistry on troponin expression which revealed significant difference between platelet-rich fibrin group compared with negative and positive controls (50.66 ± 7.2 <i>vs</i> 10.73 ± 2.39; <i>p</i> < 0.05; 50.66 ± 7.2 <i>vs</i> 26.00 ± 0.4; <i>p</i> < 0.05). <b>Conclusion:</b> Injectable platelet-rich fibrin accelerates differentiation of adipose-derived mesenchymal stem cells into cardiomyocyte-like cells.</p>","PeriodicalId":12669,"journal":{"name":"Global Cardiology Science & Practice","volume":"2024 5","pages":"e202446"},"PeriodicalIF":0.0000,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11807427/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Global Cardiology Science & Practice","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.21542/gcsp.2024.46","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Treating irreversible cardiomyocyte loss following myocardial infarction presents several therapeutic challenges. While cell therapy shows promise as a regenerative treatment for infarcted cardiac tissue, different cell sources vary in their therapeutic potential. Adipose-derived stem cells (ADSCs) have emerged as an attractive option due to their accessibility, but their limited differentiation capacity remains a significant constraint. Recent evidence suggests that injectable platelet-rich fibrin may enhance this process by stimulating the differentiation of ADSCs into cardiomyocyte-like cells. Objective: Analyse the benefit of injectable platelet-rich fibrin to accelerate the differentiation of adipose-derived mesenchymal stem cells into cardiomyocyte-like cells. Methods: This study is a true experimental randomized pos t-test design study. Adipose-derived mesenchymal stem cells were isolated from adipose tissue and expanded in culture through four passages. The characteristics of adipose-derived mesenchymal stem cells were measured by the expression of CD 34-, CD 45-, and CD 105+ using flowcytometry. The samples were divided into 3 groups, i.e., negative control (α-MEM), positive control (differentiation medium) and treatment group (platelet-rich fibrin). The assessment of GATA-4 marker expression was conducted using flowcytometry on the fifth day and troponin was conducted using immunocytochemistry on the tenth day to determine the differentiation to cardiomyocyte. Statistical analysis was performed using Student's t-tests and one-way ANOVA for data that demonstrated normal distribution as verified by the Shapiro-Wilk test. Results: Flowcytometry on GATA-4 expression revealed significant difference on addition of platelet-rich fibrin compared with negative and positive controls (68.20 ± 6.82 vs 58.15 ± 1.23; p < 0.05; 68.20 ± 6.82 vs 52.96 ± 2.02; p < 0.05). This was supported by the results of immunocytochemistry on troponin expression which revealed significant difference between platelet-rich fibrin group compared with negative and positive controls (50.66 ± 7.2 vs 10.73 ± 2.39; p < 0.05; 50.66 ± 7.2 vs 26.00 ± 0.4; p < 0.05). Conclusion: Injectable platelet-rich fibrin accelerates differentiation of adipose-derived mesenchymal stem cells into cardiomyocyte-like cells.
背景:治疗心肌梗死后不可逆的心肌细胞损失提出了几个治疗挑战。虽然细胞疗法有望作为心肌梗死组织的再生治疗,但不同细胞来源的治疗潜力各不相同。脂肪源性干细胞(ADSCs)由于其可获得性而成为一种有吸引力的选择,但其有限的分化能力仍然是一个重要的制约因素。最近的证据表明,注射富含血小板的纤维蛋白可能通过刺激ADSCs向心肌细胞样细胞的分化来促进这一过程。目的:分析注射富血小板纤维蛋白对脂肪源间充质干细胞向心肌细胞样细胞分化的促进作用。方法:本研究是一项真正的实验随机后验设计研究。从脂肪组织中分离脂肪来源的间充质干细胞,并通过四次传代培养扩增。采用流式细胞术检测cd34 -、cd45 -和cd105 +的表达,检测脂肪源性间充质干细胞的特性。将样品分为阴性对照组(α-MEM)、阳性对照组(分化培养基)和治疗组(富血小板纤维蛋白)3组。第5天采用流式细胞术检测GATA-4标记物表达,第10天采用免疫细胞化学检测肌钙蛋白,以确定向心肌细胞的分化情况。采用学生t检验和单因素方差分析对经Shapiro-Wilk检验证实为正态分布的数据进行统计分析。结果:流式细胞术检测GATA-4表达,富血小板纤维蛋白添加量与阴性对照组和阳性对照组比较差异有统计学意义(68.20±6.82 vs 58.15±1.23;P < 0.05;68.20±6.82 vs 52.96±2.02;P < 0.05)。免疫细胞化学对肌钙蛋白表达的结果也支持了这一观点,富血小板纤维蛋白组与阴性和阳性对照组相比有显著差异(50.66±7.2 vs 10.73±2.39;P < 0.05;50.66±7.2 vs 26.00±0.4;P < 0.05)。结论:注射富血小板纤维蛋白可加速脂肪源性间充质干细胞向心肌细胞样细胞的分化。