Evaluation of chondrogenesis and osteogenesis in human mesenchymal stem cells, chondrocytes, and chondroprogenitors using molecular markers, cellular markers and polarized microscopy

IF 2.6 3区生物学 Q4 CELL BIOLOGY

Differentiation Pub Date : 2025-07-25 DOI:10.1016/j.diff.2025.100898

Archa Suresh , Ayshath Ruksana C , Ganesh Parasuraman , Mohana Priya , Grace Rebekah , Christo Jeyaraj , Elizabeth Vinod

{"title":"Evaluation of chondrogenesis and osteogenesis in human mesenchymal stem cells, chondrocytes, and chondroprogenitors using molecular markers, cellular markers and polarized microscopy","authors":"Archa Suresh , Ayshath Ruksana C , Ganesh Parasuraman , Mohana Priya , Grace Rebekah , Christo Jeyaraj , Elizabeth Vinod","doi":"10.1016/j.diff.2025.100898","DOIUrl":null,"url":null,"abstract":"<div><h3>Purpose</h3><div>Fibronectin adhesion assay progenitors (FAA-CPs) and migratory assay progenitors (MCPs), subsets of mesenchymal-like stromal cells (MSCs), exhibit superior in-vitro chondrogenic potential compared to bone marrow (BM)-MSCs and chondrocytes. To assess this potential, differentiation studies followed by confirmatory staining for collagen deposition are utilized. Polarized light microscopy (PLM), based on birefringence principles, is a valuable tool for visualizing organized collagen fibers. Its use as a predictive tool for assessing chondrogenesis and osteogenesis has not been reported.</div></div><div><h3>Methods</h3><div>This study involved FAA-CPs, MCPs, chondrocytes, and BM-MSCs derived from osteoarthritic knee joints (n = 3). After phenotypic characterization, the cells underwent chondrogenic and osteogenic differentiation, followed by Picrosirius red staining and PLM analysis, including immunohistochemical analysis for collagen types I, II, and X.</div></div><div><h3>Results</h3><div>Birefringence assessment revealed greater collagen fibril alignment and significant remodeling in the BM-MSC group, which exhibited an arcade-like pattern. The MCP group displayed well-organized collagen fibrils in pericellular zones and as a peripheral band, while chondrocytes and FAA-CPs exhibited lower intensity birefringence, indicating random alignment. Areas with higher collagen type II deposition corresponded to reduced collagen type I and the absence of collagen type X, highlighting the unique fibrillar network seen with PLM was indicative of collagen type II.</div></div><div><h3>Conclusion</h3><div>While its application for osteogenesis was limited, probably due to the non-fibrillar nature of collagen type X, its value for chondrogenesis is notable. Although not directly reflecting chondrogenesis, PLM can serve as a valuable tool for gaining insights into collagen remodeling, particularly concerning collagen type II during chondrogenic differentiation.</div></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":"145 ","pages":"Article 100898"},"PeriodicalIF":2.6000,"publicationDate":"2025-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Differentiation","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0301468125000659","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Purpose

Fibronectin adhesion assay progenitors (FAA-CPs) and migratory assay progenitors (MCPs), subsets of mesenchymal-like stromal cells (MSCs), exhibit superior in-vitro chondrogenic potential compared to bone marrow (BM)-MSCs and chondrocytes. To assess this potential, differentiation studies followed by confirmatory staining for collagen deposition are utilized. Polarized light microscopy (PLM), based on birefringence principles, is a valuable tool for visualizing organized collagen fibers. Its use as a predictive tool for assessing chondrogenesis and osteogenesis has not been reported.

Methods

This study involved FAA-CPs, MCPs, chondrocytes, and BM-MSCs derived from osteoarthritic knee joints (n = 3). After phenotypic characterization, the cells underwent chondrogenic and osteogenic differentiation, followed by Picrosirius red staining and PLM analysis, including immunohistochemical analysis for collagen types I, II, and X.

Results

Birefringence assessment revealed greater collagen fibril alignment and significant remodeling in the BM-MSC group, which exhibited an arcade-like pattern. The MCP group displayed well-organized collagen fibrils in pericellular zones and as a peripheral band, while chondrocytes and FAA-CPs exhibited lower intensity birefringence, indicating random alignment. Areas with higher collagen type II deposition corresponded to reduced collagen type I and the absence of collagen type X, highlighting the unique fibrillar network seen with PLM was indicative of collagen type II.

Conclusion

While its application for osteogenesis was limited, probably due to the non-fibrillar nature of collagen type X, its value for chondrogenesis is notable. Although not directly reflecting chondrogenesis, PLM can serve as a valuable tool for gaining insights into collagen remodeling, particularly concerning collagen type II during chondrogenic differentiation.

查看原文本刊更多论文

利用分子标记、细胞标记和极化显微镜评价人间充质干细胞、软骨细胞和软骨祖细胞的软骨形成和成骨

目的：纤连蛋白粘附试验祖细胞（FAA-CPs）和迁移试验祖细胞（MCPs）是间充质样基质细胞（MSCs）的亚群，与骨髓(BM)-MSCs和软骨细胞相比，它们在体外表现出更强的成软骨潜能。为了评估这一潜力，分化研究随后证实染色胶原沉积被利用。偏振光显微镜（PLM），基于双折射原理，是一个有价值的工具，可视化组织胶原纤维。它作为评估软骨形成和成骨形成的预测工具尚未报道。方法本研究涉及骨关节炎膝关节的fa - cps、MCPs、软骨细胞和BM-MSCs （n = 3）。表型鉴定后，细胞进行软骨和成骨分化，随后进行Picrosirius红染色和PLM分析，包括I型、II型和x型胶原的免疫组织化学分析。结果双折射评估显示，BM-MSC组胶原纤维排列更大，重构明显，呈拱廊状。MCP组在细胞周带和外周带中显示组织良好的胶原原纤维，而软骨细胞和fa - cps表现出较低强度的双折射，表明随机排列。II型胶原沉积较多的区域对应于I型胶原减少和X型胶原缺失，突出显示PLM中独特的纤维网络，表明II型胶原。结论X型胶原的非纤原性可能限制了其在成骨方面的应用，但其在软骨形成方面的价值是显著的。虽然PLM不能直接反映软骨形成，但它可以作为一种有价值的工具，用于深入了解胶原重塑，特别是软骨形成分化过程中的II型胶原。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Differentiation 生物-发育生物学

CiteScore

4.10

自引率

3.40%

发文量

审稿时长

51 days

期刊介绍： Differentiation is a multidisciplinary journal dealing with topics relating to cell differentiation, development, cellular structure and function, and cancer. Differentiation of eukaryotes at the molecular level and the use of transgenic and targeted mutagenesis approaches to problems of differentiation are of particular interest to the journal. The journal will publish full-length articles containing original work in any of these areas. We will also publish reviews and commentaries on topics of current interest. The principal subject areas the journal covers are: • embryonic patterning and organogenesis • human development and congenital malformation • mechanisms of cell lineage commitment • tissue homeostasis and oncogenic transformation • establishment of cellular polarity • stem cell differentiation • cell reprogramming mechanisms • stability of the differentiated state • cell and tissue interactions in vivo and in vitro • signal transduction pathways in development and differentiation • carcinogenesis and cancer • mechanisms involved in cell growth and division especially relating to cancer • differentiation in regeneration and ageing • therapeutic applications of differentiation processes.