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. 2022 Jan 25;40(1):14-21.
doi: 10.7518/hxkq.2022.01.003.

Effect of growth differentiation factor 11 on the steatosis of condylar chondrocytes in temporomandibular joint osteoarthritic mice

[Article in English, Chinese]
Ruoxin Wang  1   2 , Qian Liu  2 , Feng He  2 , Mian Zhang  2 , Helin Wang  3
Affiliations

Effect of growth differentiation factor 11 on the steatosis of condylar chondrocytes in temporomandibular joint osteoarthritic mice

[Article in English, Chinese]
Ruoxin Wang et al. Hua Xi Kou Qiang Yi Xue Za Zhi. .

Abstract
in English, Chinese

Objectives: To observe the effect of growth differentiation factor 11 (GDF11) on the steatosis of condylar chondrocytes in temporomandibular joint (TMJ) osteoarthritic mice.

Methods: Six-week-old female C57 mice were stimulated with unilateral anterior crossbite (UAC) and locally injected with GDF11. TMJ sections were stained with safranin O, and GDF11 and Adiponectin in the TMJ cartilage were detected by immunohistochemical staining. GDF11, Adiponectin, type Ⅱ collagen (Col-Ⅱ), Aggrecan, type Ⅰ collagen (Col-Ⅰ), fatty acid-binding protein 4 (FABP4), and Perilipin 1 mRNA were detected by real-time polymerase chain reaction (real-time PCR).

Results: The thickness and safranin O-positive area of TMJ cartilage in the UAC groups, as well as the mRNA expression of Col-Ⅱ and aggrecan, significantly decreased. The ratio of Adiponectin-positive cells significantly increased. The mRNA expression levels of Col-Ⅰ, Adiponectin, FABP4, and Perilipin 1 significantly increased. The TMJ cartilage of the GDF11 injection groups significantly thickened, and the safranin O-positive area significantly increased. The mRNA expression of Col-Ⅱ and Aggrecan significantly increased. The ratio of Adiponectin-positive cells significantly decreased. The mRNA expression levels of Col-I, Adiponectin, FABP4, and Perilipin 1 significantly decreased.

Conclusions: Extensive steatosis of chondrocytes and decreased expression of GDF11 in the degenerated cartilage of TMJ osteoarthritis are induced by UAC stimulation. Supplementation of exogenous GDF11 can effectively inhibit the steatosis of chondrocytes.

目的: 观察生长分化因子11（GDF11）对小鼠颞下颌关节（TMJ）骨关节炎（OA）髁突软骨细胞脂肪化的影响。方法: 对雌性6周龄C57小鼠施加单侧前牙反牙合（UAC）刺激并局部注射GDF11。取TMJ髁突软骨进行番红O染色；GDF11、脂联素（Adiponectin）免疫组织化学染色；实时定量聚合酶链式反应（real-time PCR）检测GDF11、Adiponectin、Ⅱ型胶原（Col-Ⅱ）、Aggrecan、Ⅰ型胶原（Col-Ⅰ）、脂肪酸结合蛋4（FABP4）、脂滴包被蛋白1（Perilipin 1）的mRNA含量。结果: UAC实验组髁突软骨显著变薄、番红O阳性面积显著减少；Col-Ⅱ和Aggrecan的mRNA表达量显著降低；Adiponectin阳性细胞率显著增加；Col-Ⅰ、Adiponectin、FABP4、Perilipin 1的mRNA表达量显著上升。GDF11注射后髁突软骨显著增厚、番红O阳性面积显著增加；Col-Ⅱ和Aggrecan的mRNA表达量显著升高；Adiponectin阳性细胞率显著减少；Col-Ⅰ、Adiponectin、FABP4、Perilipin 1的mRNA表达量显著降低。结论: UAC刺激诱导的髁突退变软骨中GDF11表达降低并存在软骨细胞脂肪化，补充外源性GDF11可有效抑制软骨细胞脂肪化。.

Keywords: chondrocytes; growth differentiation factor 11; osteoarthritis; steatosis; temporomandibular joint.

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Conflict of interest statement

利益冲突声明：作者声明本文无利益冲突。

Figures

图 1
图 1. 正常小鼠和TMJ OA小鼠髁突软骨番红O染色结果 番红O染色 × 200
Fig 1 Safranin O staining of TMJ condylar cartilage in normal and TMJ OA mice safranin O staining × 200
图 2
图 2. 正常小鼠和TMJ OA小鼠髁突软骨退变情况对比
Fig 2 Comparison of TMJ condylar cartilage in normal and TMJ OA mice A：软骨厚度；B：番红O阳性面积；C：Col-Ⅱ mRNA表达量；D：Aggrecan mRNA表达量；E：Col-Ⅰ表达量。a：3周对照组；b：3周UAC实验组；c：7周对照组；d：7周UAC实验组；e：11周对照组；f：11周UAC实验组。与同期对照组相比，*P<0.05，**P<0.01。
图 3
图 3. 正常小鼠和TMJ OA小鼠髁突软骨Adiponectin免疫组织化学染色结果 免疫组织化学染色 × 200
Fig 3 Adiponectin immunohistochemical staining of TMJ condylar cartilage in normal and TMJ OA mice immunohistochemical staining × 200
图 4
图 4. 正常小鼠和TMJ OA小鼠髁突软骨内脂肪化情况对比
Fig 4 Comparison of steatosis in TMJ condylar cartilage between normal and TMJ OA mice A：Adiponectin阳性细胞率；B：Adiponectin mRNA表达量；C：FABP4 mRNA表达量；D：Perilipin 1 mRNA表达量。a：3周对照组；b：3周UAC实验组；c：7周对照组；d：7周UAC实验组；e：11周对照组；f：11周UAC实验组。与同期对照组相比，**P<0.01。
图 5
图 5. 正常小鼠和TMJ OA小鼠髁突软骨GDF11免疫组织化学染色结果 免疫组织化学染色 × 200
Fig 5 GDF11 immunohistochemical staining of TMJ condylar cartilage in normal and TMJ OA mice immunohistochemical staining × 200
图 6
图 6. 正常小鼠和TMJ OA小鼠髁突软骨内GDF11的表达对比
Fig 6 Comparison of GDF11 expression in TMJ condylar cartilage between normal and TMJ OA mice A：GDF11阳性细胞率；B：GDF11 mRNA表达量。a：3周对照组；b：3周UAC实验组；c：7周对照组；d：7周UAC实验组；e：11周对照组；f：11周UAC实验组。与同期对照组相比，*P<0.05；**P<0.01。
图 7
图 7. PBS注射和GDF11注射小鼠髁突软骨退变、脂肪化情况对比
Fig 7 Comparison of degeneration and steatosis in TMJ condylar cartilage between mice with PBS or GDF11 injection A：髁突软骨 番红O染色 × 200；B：软骨厚度；C：番红O阳性面积；D：髁突软骨Adiponectin 免疫组织化学染色 × 200；E：Adiponectin阳性细胞率；F：Adiponectin mRNA表达量。a：7周UAC+PBS注射组；b：7周UAC+GDF11注射组；c：11周UAC+PBS注射组；d：11周UAC+GDF11注射组。与同期对照组相比，*P<0.05；**P<0.01。
图 8
图 8. PBS注射和GDF11注射小鼠髁突软骨退变、脂肪化相关分子表达对比
Fig 8 Comparison of molecular expression related to degeneration and steatosis in TMJ condylar cartilage between mice with PBS or GDF11 injection A：Col-Ⅱ mRNA表达量；B：Aggrecan mRNA表达量；C：Col-Ⅰ表达量；D：FABP4 mRNA表达量；E：Perilipin 1 mRNA表达量。a：7周UAC+PBS注射组；b：7周UAC+GDF11注射组；c：11周UAC+PBS注射组；d：11周UAC+GDF11注射组。与同期对照组相比，**P<0.01。
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