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| Effect of the new hydrogels loaded with plateletrich fibrin on chondrogenic differentiation of BMSCs |
| CHEN Xing-guang1,2, ZOU Cheng-da1,2, SU Guang-hao1, ZHANG Ya1, LU Min-hua3, WANG Xiao-dong2 |
| （1. Pediatrics Research Institute， 2. Department of Orthopaedics, Children′s Hospital of Soochow University, Suzhou Jiangsu 215025; 3. Department of Orthopaedics, Suzhou Dushuhu Public Hospital, Suzhou Jiangsu 215025, China） |
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Abstract Objective: To explore effect of the new hydrogels loaded with plateletrich fibrin(PRF) on chondrogenic differentiation of BMSCs. Methods: 2% chitosan (CS) solution, 56% βglycerophosphate (βGP) solution and 0.1% genipin solution were mixed with the ratio of 6 ∶1 ∶1 to get the CS/GP/0.1% genipin mixed solution, and measured its gelformation time at 37 ℃.Then PRF was prepared by once centrifugation (1 200 r/min, 12 min), and the freezedrying PRF was ground into powder, which was added into that mixed solution. New type hydrogels were made with PRF at 37 ℃. The BMSCs were seeded on the surface of the hydrogels, and the state of cell growth was observed by FDA fluorescent staining. For chondrogenic induction, BMSCs were then cultured in the 24well plate, by adding the new type of hydrogels, chitosan thermosensitive hydrogels or empty control at the same time. Effect of the new type of hydrogels on chondrogenic differentiation were observed by HE staining, safranin Ofast green staining, alcian blue staining and immunohistochemical staining of type Ⅱ collagen 2 weeks after cell culture. The amount of glycosaminoglycans (GAGs) in the supernatant was quantitatively determind by alcian blue colorimetric assay 4 weeks after cell culture. The amount of type Ⅱ collagen in supernatant was quantitated by ELISA 4 weeks after cell culture. Results: The gelformation time of the volume ratio of 6 ∶1 ∶1 of the CS/GP/0.1% genipin mixed solution was about 8 min at 37℃. The BMSCs seeded on the surface of the hydrogels presented good proliferation. For the group of the new type of hydrogels, the result of HE staining, safranin Ofast green staining, alcian blue staining and immunohistochemical staining of type Ⅱ collagen were positive, and the other two group were weakly positive. There are GAGs and type Ⅱ collagen in the supernatant of three groups. But there are statistical significance between the group of the new type of hydrogel and the other groups for the difference in secretion of GAGs and type Ⅱ collagen (P<0.05). Conclusion: That hydrogel have a good biocompatibility with BMSCs, and the BMSCs seeded on the surface of the hydrogels presented good proliferation. The new type hydrogels with PRF were able to promote chondrogenic differentiation to a certain degree.
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Received: 23 March 2018
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2018, Vol. 28 
