Background and ObjectivesOne of the most cellular source used for cartilage tissue engineering are mesenchymal stem cells (MSCs). In present study, human MSCs were used as cellular source. Since scaffold plays an important role in tissue engineering the aim of this study is to assess fibrin scaffold ability in chondrogenic differentiation of adipose-derived mesenchymal stem cells (ADMSCs).MethodsADMSCs were isolated and cultured in DMEM medium supplemented with 10% FBS. Also ADMSCs expanded and characterised by flow cytometry. ADMSCs expressed CD44, CD90, CD105 but not CD34. After trypsinization, cells were entered within the fibrin scaffold. Then, chondrogenic medium was added to the scaffold. Seven days after cell culture, cell viability and proliferation were assessed by MTT test. Finally, 14 days after the ending of chondrogenic differentiation, analysis of chondrogenic genes expression was evaluated by RT-PCR and Real time PCR. Also, formation and development of chondrocyte cells was analysed by histological and immunohistochemistry evaluations.ResultsViability and proliferation as well as chondrogenic genes expression within fibrin scaffold increased significantly compared with control group (cells free scaffold). Also, histological and immunohistochemistry evaluation showed that chondrocyte cells and collagen type II are formed on fibrin scaffold.ConclusionsFibrin is a suitable scaffold for chondrogenic differentiation of ADMSCs.

This study presents a comparative assessment of adipose-derived stem cell (ADSCs) proliferation rates and their viability on five different scaffolds. Five different biomaterial scaffolds were prepared: alginate, poly lactic-co-glycolic acid, fibrin glue, inactive platelet-rich plasma, and active plateletrich plasma (APRP). Stem cells were isolated from human adipose tissue. Flow cytometry analysis was performed. Specifically, adipogenesis/osteogenesis/chondrogenesis-associated genes expression was analyzed by real-time polymerase chain reaction. These cells were seeded in the prepared scaffolds. After 14 days, the proliferation and viability of MSCs were evaluated using an MTT assay. Also, stemness genes expression was analyzed with the reverse transcriptasepolymerase chain reaction (RT-PCR) method. In addition, the DNA content assay was also performed. The obtained results showed a significant difference between cell proliferation and viability of different scaffolds. APRP and alginate were shown to be the most and least suitable scaffolds in terms of enhancing cell proliferation and maintaining cell viability respectively (p < .05). RT-PCR reactions demonstrated the expression of the various stemness-related markers (Nanog, Octamer4A, and Sox2) when ADSC cells were grown separately on the five different scaffolds. Our study indicates that compared with the scaffolds, APRP could be the best scaffold for support of ADSC proliferation. ARTICLE HISTORY

Background:Although progenitor cells have been observed in articular cartilage, this part has a limited ability to repair due to a lack of blood supply. Formerly, tissue engineering was mainly based on collecting chondrocytes from the joint surface, culturing them on resorbable scaffolds such as poly D, L-lactic glycolic acid (PLGA) and then autologous transplantation. In recent times, due to difficulties in collecting chondrocytes, most of the researchers are focused on stem cells for producing these cells. Among the important factors in this approach, is using appropriate scaffolds with good mechanical and biological properties to provide optimal environment for growth and development of stem cells. In this study, we evaluated the potential of fibrin glue, PLGA and alginate scaffolds in providing a suitable environment for growth and chondrogenic differentiation of mesenchymal stem cells (MSCs) in the presence of transforming growth factor-β3.Materials and Methods:Fibrin glue, PLGA and alginate scaffolds were prepared and MSCs were isolated from human adipose tissue. Cells were cultured separately on the scaffolds and 2 weeks after differentiation, chondrogenic genes, cell proliferation ability and morphology in each scaffold were evaluated using real time-polymerase chain reaction, MTT chondrogenic assay and histological examination, respectively.Results:Proliferation of differentiated adipose tissue derived mesenchymal stem cells (AD-MSCs) to chondrogenic cells in Fibrin glue were significantly higher than in other scaffolds. Also, Fibrin glue caused the highest expression of chondrogenic genes compared to the other scaffolds. Histological examination revealed that the pores of the Fibrin glue scaffolds were filled with cells uniformly distributed.Conclusion:According to the results of the study, it can be concluded that natural scaffolds such as fibrin can be used as an appropriate environment for cartilage differentiation.

ABSTRACT:The ability of cartilage to repair damage is limited due to lack of blood vessels and low cell density. Recently, tissue engineering is considerably preferred to other treatments as a way to solve this problem. Regardless of cell sources, one of the crucial factors in tissue engineering is to select an appropriate scaffold, which is essential for healing and renewal procedure of tissues in vivo and in vitro. Mesenchymal stem cells (MSCs) were isolated from adipose tissue in liposuction surgeries by use of collagenase enzyme. After verification by flow cytometry methods, adipose-derived mesenchymal stem cells (ADMSCs) were embedded into alginate and agarose scaffolds, separately; and then they were cultured in chondrogenic medium for 3 weeks. The ability of alginate and agarose scaffolds was assessed by use of MTT assay and histological analysis. In addition, analysis of chondrogenic genes expression by Real-time PCR was done. The obtained data were analyzed statistically by means of SPSS software. There was a significant difference between alginate and agarose groups in maintaining cells viable but, about chondrogenic differentiation analyzed by use of real-time PCR, statistical analysis has shown a significant difference in expression of aggrecan (as a chondrocyte-specific gene) and collagen II (as an chondrocyte-specific gene) between cell/alginate and cell/agarose and MSCs (p<0.05). Chondrocyte differentiation of cells was verified by histological analysis. Alginate scaffold can provide a suitable environment for chondrogenic differentiation of adipose derived mesenchymal stem cells.

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