Evaluation of osteoconductive effect of polycaprolactone (PCL) scaffold treated with Aloe vera on adipose-derived mesenchymal stem cells (ADSCs).

IF 1.5 Q4 CELL BIOLOGY American journal of stem cells Pub Date : 2023-10-20 eCollection Date: 2023-01-01
Maryam Teymori, Ehsan Karimi, Ehsan Saburi
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Abstract

Background: Adipose-derived mesenchymal stem cells (ADSCs) hold promise for bone tissue engineering because of their ability to differentiate into a variety of cell lineages. In tissue engineering, composite scaffolds made of natural and synthetic polymers have also attracted interest. Modification of scaffolds with various substances, including Aloe Vera, is expected to play a useful role in the repair of damaged tissues, including bone.

Method: ADSCs were isolated and seeded in three groups on an Aloe Vera-modified PCL scaffold: 1. Polycaprolactone (PCL) scaffold group, 2. PCL/Aloe Vera scaffold group, and 3. TCPS (Tissue Culture Polystyrene) group. Subsequently, staining with Oil red and Alizarin Red was performed to assess the ability of ADSCs to differentiate into fat and bone cells. Cell viability was determined by the resazurin assay on days 1, 3, and 5. Calcium content and alkaline phosphatase activity (ALP) were determined with kits on days 7, 14, and 21. RNA was extracted, and cDNA was synthesized. Finally, the expression of marker genes for bone differentiation like osteogenic markers such as Osteonectin (ON), Osteocalcin (OC), RUNX Family Transcription Factor 2 (RUNX2), Collagen type I alpha 1 (COL1) was evaluated by real-time PCR.

Results: Aloe vera-treated PCL scaffolds showed improved biocompatibility compared with untreated scaffolds (P<0.05). In addition, treated scaffolds promoted osteogenic differentiation of ADSCs, as evidenced by increased expression of osteogenic markers such ON, OC, RUNX2, COL1 compared with PCL scaffold and TCPS (P<0.05). Furthermore, ALP and calcium content assay confirmed improved mineral deposition on PCL scaffolds treated with Aloe vera, indicating enhanced osteoconductivity (P<0.05).

Conclusion: Our data suggest that a PCL scaffold mixed with Aloe Vera gel has promising osteoconductive potential, which can be used as a natural polymer for tissue engineering of bone and promote bone regeneration.

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芦荟处理聚己内酯(PCL)支架对脂肪源性间充质干细胞(ADSCs)的骨传导效果评价。
背景:脂肪源性间充质干细胞(ADSCs)具有分化成多种细胞系的能力,因此在骨组织工程中具有应用前景。在组织工程中,由天然和合成聚合物制成的复合支架也引起了人们的兴趣。用包括芦荟在内的各种物质修饰支架,有望在包括骨在内的受损组织的修复中发挥有用的作用。方法:分离ADSCs并将其植入芦荟修饰的PCL支架上,分为三组:聚己内酯(PCL)支架组;PCL/芦荟支架组;TCPS(组织培养聚苯乙烯)组。随后,采用油红和茜素红染色来评估ADSCs向脂肪细胞和骨细胞分化的能力。在第1、3、5天用瑞唑脲法测定细胞活力。分别于第7、14、21天用试剂盒测定各组钙含量和碱性磷酸酶活性(ALP)。提取RNA,合成cDNA。最后,通过实时荧光定量PCR检测骨分化标志物基因如骨连接素(ON)、骨钙素(OC)、RUNX家族转录因子2 (RUNX2)、I型胶原α 1 (COL1)等的表达情况。结果:经芦荟处理的PCL支架的生物相容性较未处理的PCL支架有明显改善(PON、OC、RUNX2、COL1较PCL支架和TCPS)。结论:经芦荟凝胶处理的PCL支架具有良好的骨传导潜力,可作为骨组织工程的天然高分子材料,促进骨再生。
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