In vivo cartilage formation using chondrogenic-differentiated human adipose-derived mesenchymal stem cells mixed with fibrin glue.
Human adipose-derived mesenchymal stem cells (MSCs) were differentiated into chondrogenic MSCs, and fibrin glue was used together to explore the feasibility of whether cartilages can be generated in vivo by injecting the differentiated cells. Mesenchymal stem cells extracted from human adipose were differentiated into chondrogenic MSCs, and such differentiated cells mixed with fibrin glue were injected subcutaneously into the back of the nude mouse. In addition to visual evaluation of the tissues formed after 4, 8, and 12 weeks, hematoxylin-eosin staining, Masson trichrome staining, measurement of glycosaminoglycan concentration using dimethylmethylene blue, agreecan through reverse transcriptase-polymerase chain reaction, type II collagen, and expression of SOX-9 were verified. Moreover, the results were compared with 2 groups of controls: 1 control group that received only injection of chondrogenic-differentiated MSC and the supporting control group that received only fibrin glue injection. For the experimental group, cartilage-like tissues were formed after 4, 8, and 12 weeks. Formation of cartilage tissues was not observed in any of 4, 8, and 12 weeks of the control group. The supporting control group had only a small structure formation after 4 weeks, but the formed structure was completely decomposed by the 8th and 12th weeks. The range of staining dramatically increased with time at 4, 8, and 12 weeks in Masson trichrome staining. The concentration of glycosaminoglycan also increased with time. The increased level was statistically significant with more than 3 times more after 8 weeks compared with 4 weeks and more than 2 times more after 12 weeks compared with 8 weeks. Also, in reverse transcriptase-polymerase chain reaction at 4, 8, and 12 weeks, all results expressed a cartilage-specific gene called aggrecan, type II collagen, and SOX-9. The study verified that the chondrogenic-differentiated MSCs derived from human adipose tissues with fibrin glue can proliferate and form new cartilage. Our findings suggest that formation of cartilages in vivo is possible.
Jung
Sung-No
SN
From the Department of Plastic Surgery, College of Medicine, Catholic University of Korea, Seoul, Korea.
Rhie
Jong Won
JW
Kwon
Ho
H
Jun
Young Joon
YJ
Seo
Je-Won
JW
Yoo
Gyeol
G
Oh
Deuk Young
DY
Ahn
Sang Tae
ST
Woo
Jihyoun
J
Oh
Jieun
J
D017667:Adipocytes / Q000502:physiology*; D053674:Aggrecans / Q000032:analysis; D000423:Alcian Blue; D000818:Animals; D001391:Azo Compounds; D002356:Cartilage / Q000033:anatomy & histology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D020219:Chondrogenesis / Q000502:physiology*; D024043:Collagen Type II / Q000032:analysis; D004396:Coloring Agents; D004801:Eosine Yellowish-(YS); D005240:Feasibility Studies; D005260:Female; D015718:Fibrin Tissue Adhesive / Q000627:therapeutic use*; D006025:Glycosaminoglycans / Q000032:analysis; D006801:Humans; D007279:Injections, Subcutaneous; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008739:Methyl Green; D008751:Methylene Blue / Q000031:analogs & derivatives; D051379:Mice; D008819:Mice, Nude; D010619:Phenazines; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000032:analysis; D013997:Time Factors
D053674:Aggrecans; D001391:Azo Compounds; D024043:Collagen Type II; D004396:Coloring Agents; D015718:Fibrin Tissue Adhesive; D006025:Glycosaminoglycans; D010619:Phenazines; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; C435946:dimethylmethylene blue; C017768:trichrome stain; D008739:Methyl Green; D000423:Alcian Blue; D008751:Methylene Blue; D004801:Eosine Yellowish-(YS); C009195:safranine T