MODEL ESTABLISHING OF PARTIAL-THICKNESS ARTICULAR CARTILAGE INJURY AND RELATIONSHIPS BETWEEN ACTIVATION OF CELLS AND EXPRESSION OF INTEGRIN β<sub>1</sub> IN A RAT MODEL_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANGKaibin , SHIJing , LIYang , JIANGYiqiu , DINGPeng , YANGXiaofei ,  GUIJianchao

Department of Orthopedics, Nanjing Hospital Affiliated to Nanjing Medical University, Nanjing Jiangsu, 210000, P. R. China;

Corresponding author：

GUIJianchao, Email: gui1997@126.com

Keywords：

Partial-thickness articular cartilage injury; Cartilage progenitor cells; CD105; BrdU; Integrin β₁; Rat

DOI：

10.7507/1002-1892.20160087

Video：

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Objective To investigate the relationships between the expression of integrin β₁ and activated cells in a partial-thickness articular cartilage injury model of adult rats. Method Forty-five male Sprague Dawley rats (aged 10 weeks and weighing 300-400 g) were randomly divided into operated group (n=15) , sham-operated group (n=15) , and control group (n=15) . Partial-thickness articular cartilage injury model was made by scarification in operated group, direct suture after opening of the knee joint was performed in sham-operated group, and no operation was done in control group. Five rats were sacrificed at 1, 7, and 14 days after operation respectively for macroscopic evaluation, HE staining, Safranin O staining, CD105, BrdU, CD105/integrin β₁ immunofluorescence and double labeling staining. The histological score of HE staining, gray value of Safranin O staining and CD105-positive cells count were compared among groups at each time point. Results Macroscopic evaluation showed chondromalacia and cartilage fibrosis around the linear injury with aggravating tendency with time in operated group, but no chondromalacia and cartilage fibrosis in sham-operated and control groups. HE staining demonstrated a number of activated cells accumulating around the linear injury with nonuniform distribution in operated group, and uniform size and distribution in sham-operated and control groups. The histological scores at each time point in operated group were significantly higher than those in sham-operated group and control group (P<0.05) , but no significant difference was found between different time points in 3 groups (P>0.05) . Safranin O staining was nonuniform with hypochromasia around linear injury in operated group, but the staining was uniform in sham-operated group and control group. Gray value of Safranin O staining had no significant difference among groups and among different time points in the same group (P>0.05) . BrdU-positive and CD105-positive cells distributed unevenly around the linear injury in operated group, uniform distribution was observed in sham-operated group and control group. CD105-positive cells count in operated group was significantly higher than those in sham-operated group and control group at each time point (P<0.05) ; CD105-positive cells increased significantly with time in operated group (P<0.05) . CD105/integrinβ₁-positive cells were observed around the linear injury in operated group, but was not observed in sham-operated group and control group. Conclusions The partial-thickness articular cartilage injury model is successfully established in rats, and cartilage injury could not be repaired completely in the model. The activated cells aggregation around the linear injury can be observed, but there is no obvious relationships between activated cells and cartilage matrix. These activated cells are in proliferation and could express both CD105 and integrin β₁.

Citation： ZHANGKaibin, SHIJing, LIYang, JIANGYiqiu, DINGPeng, YANGXiaofei, GUIJianchao. MODEL ESTABLISHING OF PARTIAL-THICKNESS ARTICULAR CARTILAGE INJURY AND RELATIONSHIPS BETWEEN ACTIVATION OF CELLS AND EXPRESSION OF INTEGRIN β₁ IN A RAT MODEL. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(4): 432-439. doi: 10.7507/1002-1892.20160087 Copy

1.	Gottschling S, Saffrich R, Seckinger A, et al. Human mesenchymal stromal cells regulate initial self-renewing divisions of hematopoietic progenitor cells by a β₁-integrin-dependent mechanism. Stem Cells, 2007, 25(3) :798-806.
2.	Muhammad H, Schminke B, Miosge N. Current concepts in stem cell therapy for articular cartilage repair. Expert Opin Biol Ther, 2013, 13(4) :541-548.
3.	Dowthwaite GP, Bishop JC, Redman SN, et al. The surface of articular cartilage contains a progenitor cell population. J Cell Sci, 2004, 117(Pt 6) :889-897.
4.	Mukoyama S, Sasho T, Akatsu Y, et al. Spontaneous repair of partial thickness linear cartilage injuries in immature rats. Cell Tissue Res, 2015, 359(2) :513-520.
5.	Hunziker EB. Growth-factor-induced healing of partial-thickness defects in adult articular cartilage. Osteoarthritis Cartilage, 2001, 9(1) :22-32.
6.	Namba RS, Meuli M, Sullivan KM, et al. Spontaneous repair of superficial defects in articular cartilage in a fetal lamb model. J Bone Joint Surg (Am), 1998, 80(1) :4-10.
7.	Baptista LS, Silva KR, Pedrosa CS, et al. Bioengineered cartilage in a scaffold-free method by human cartilage-derived progenitor cells:a comparison with human adipose-derived mesenchymal stromal cells. Artif Organs, 2013, 37(12) :1068-1075.
8.	Sandell LJ. Etiology of osteoarthritis:genetics and synovial joint development. Nat Rev Rheumatol, 2012, 8(2) :77-89.
9.	Tsuruoka H, Sasho T, Yamaguchi S, et al. Maturation-dependent spontaneous healing of partial thickness cartilage defects in infantile rats. Cell Tissue Res, 2011, 346(2) :263-271.
10.	Yoshioka M, Kubo T, Coutts RD, et al. Differences in the repair process of longitudinal and transverse injuries of cartilage in the rat knee. Osteoarthritis Cartilage, 1998, 6(1) :66-75.
11.	Seol D, McCabe DJ, Choe H, et al. Chondrogenic progenitor cells respond to cartilage injury. Arthritis Rheum, 2012, 64(11) :3626-3637.
12.	Kaneshiro N, Sato M, Ishihara M, et al. Bioengineered chondrocyte sheets may be potentially useful for the treatment of partial thickness defects of articular cartilage. Biochem Biophys Res Commun, 2006, 349(2) :723-731.
13.	Goldring MB. Chondrogenesis, chondrocyte differentiation, and articular cartilage metabolism in health and osteoarthritis. Ther Adv Musculoskel Dis, 2012, 4(4) :269-285.
14.	王敏, Peter Chen,刘超, 等. 早期兔骨关节炎软骨CD105+/CD166+细胞及其软骨分化潜能的实验研究. 中国修复重建外科杂志, 2013, 27(7) :793-799.
15.	Yu YP, Wang Q, Liu YC, et al. Molecular basis for the targeted binding of RGD-containing peptide to integrin αVβ3. Biomaterials, 2014, 35(5) :1667-1675.
16.	Zwolanek D, Flicker M, Kirstätter E, et al. β₁ integrins mediate attachment of mesenchymal stem cells to cartilage lesions. Biores Open Access, 2015, 4(1) :39-53.
17.	闫德雄, 苏踊跃, 唐红英, 等. 整合素β₁在人表皮细胞株HaCaT增殖中影响的研究. 第三军医大学学报, 2006, 28(8) :760-762.
18.	Joos H, Wildner A, Hogrefe C, et al. Interleukin-1 beta and tumor necrosis factor alpha inhibit migration activity of chondrogenic progenitor cells from non-fibrillated osteoarthritic cartilage. Arthritis Res Ther, 2013, 15(5) :R119.
19.	Baptista LS, Silva KR, Pedrosa CS, et al. Bioengineered cartilage in a scaffold-free method by human cartilage-derived progenitor cells:a comparison with human adipose-derived mesenchymal stromal cells. Artif Organs, 2013, 37(12) :1068-1075.
20.	张向鑫, 马瑞雪, 李天友, 等. 整合素β₁在髋关节发育不良髋臼软骨退变中的作用. 临床小儿外科杂志, 2009, 8(3) :29-33.
21.	钟清玲, 刘德伍, 刘繁荣, 等. 糖尿病大鼠表皮干细胞及其增殖分化相关蛋白的研究. 中国修复重建外科杂志, 2010, 24(2) :133-137.
22.	Jansen EJ, Emans PJ, Van Rhijn LW, et al. Development of partial-thickness articular cartilage injury in a rabbit model. Clin Orthop Relat Res, 2008, 466(2) :487-494.
23.	王凡, 李爱冬, 羊惠君, 等. 转化生长因子-β对骺软骨细胞增殖和PCNA表达的影响. 华西医大学报, 2000, 31(4) :460-462, 467.
24.	任戈亮, 桂鉴超, 王黎明, 等. Ⅱ型胶原羧基端端肽对骨关节炎早期诊断和病情评估的实验研究. 南京医科大学学报(自然科学版), 2010, 30(5) :653-657.

1. Gottschling S, Saffrich R, Seckinger A, et al. Human mesenchymal stromal cells regulate initial self-renewing divisions of hematopoietic progenitor cells by a β₁-integrin-dependent mechanism. Stem Cells, 2007, 25(3) :798-806.
2. Muhammad H, Schminke B, Miosge N. Current concepts in stem cell therapy for articular cartilage repair. Expert Opin Biol Ther, 2013, 13(4) :541-548.
3. Dowthwaite GP, Bishop JC, Redman SN, et al. The surface of articular cartilage contains a progenitor cell population. J Cell Sci, 2004, 117(Pt 6) :889-897.
4. Mukoyama S, Sasho T, Akatsu Y, et al. Spontaneous repair of partial thickness linear cartilage injuries in immature rats. Cell Tissue Res, 2015, 359(2) :513-520.
5. Hunziker EB. Growth-factor-induced healing of partial-thickness defects in adult articular cartilage. Osteoarthritis Cartilage, 2001, 9(1) :22-32.
6. Namba RS, Meuli M, Sullivan KM, et al. Spontaneous repair of superficial defects in articular cartilage in a fetal lamb model. J Bone Joint Surg (Am), 1998, 80(1) :4-10.
7. Baptista LS, Silva KR, Pedrosa CS, et al. Bioengineered cartilage in a scaffold-free method by human cartilage-derived progenitor cells:a comparison with human adipose-derived mesenchymal stromal cells. Artif Organs, 2013, 37(12) :1068-1075.
8. Sandell LJ. Etiology of osteoarthritis:genetics and synovial joint development. Nat Rev Rheumatol, 2012, 8(2) :77-89.
9. Tsuruoka H, Sasho T, Yamaguchi S, et al. Maturation-dependent spontaneous healing of partial thickness cartilage defects in infantile rats. Cell Tissue Res, 2011, 346(2) :263-271.
10. Yoshioka M, Kubo T, Coutts RD, et al. Differences in the repair process of longitudinal and transverse injuries of cartilage in the rat knee. Osteoarthritis Cartilage, 1998, 6(1) :66-75.
11. Seol D, McCabe DJ, Choe H, et al. Chondrogenic progenitor cells respond to cartilage injury. Arthritis Rheum, 2012, 64(11) :3626-3637.
12. Kaneshiro N, Sato M, Ishihara M, et al. Bioengineered chondrocyte sheets may be potentially useful for the treatment of partial thickness defects of articular cartilage. Biochem Biophys Res Commun, 2006, 349(2) :723-731.
13. Goldring MB. Chondrogenesis, chondrocyte differentiation, and articular cartilage metabolism in health and osteoarthritis. Ther Adv Musculoskel Dis, 2012, 4(4) :269-285.
14. 王敏, Peter Chen,刘超, 等. 早期兔骨关节炎软骨CD105+/CD166+细胞及其软骨分化潜能的实验研究. 中国修复重建外科杂志, 2013, 27(7) :793-799.
15. Yu YP, Wang Q, Liu YC, et al. Molecular basis for the targeted binding of RGD-containing peptide to integrin αVβ3. Biomaterials, 2014, 35(5) :1667-1675.
16. Zwolanek D, Flicker M, Kirstätter E, et al. β₁ integrins mediate attachment of mesenchymal stem cells to cartilage lesions. Biores Open Access, 2015, 4(1) :39-53.
17. 闫德雄, 苏踊跃, 唐红英, 等. 整合素β₁在人表皮细胞株HaCaT增殖中影响的研究. 第三军医大学学报, 2006, 28(8) :760-762.
18. Joos H, Wildner A, Hogrefe C, et al. Interleukin-1 beta and tumor necrosis factor alpha inhibit migration activity of chondrogenic progenitor cells from non-fibrillated osteoarthritic cartilage. Arthritis Res Ther, 2013, 15(5) :R119.
19. Baptista LS, Silva KR, Pedrosa CS, et al. Bioengineered cartilage in a scaffold-free method by human cartilage-derived progenitor cells:a comparison with human adipose-derived mesenchymal stromal cells. Artif Organs, 2013, 37(12) :1068-1075.
20. 张向鑫, 马瑞雪, 李天友, 等. 整合素β₁在髋关节发育不良髋臼软骨退变中的作用. 临床小儿外科杂志, 2009, 8(3) :29-33.
21. 钟清玲, 刘德伍, 刘繁荣, 等. 糖尿病大鼠表皮干细胞及其增殖分化相关蛋白的研究. 中国修复重建外科杂志, 2010, 24(2) :133-137.
22. Jansen EJ, Emans PJ, Van Rhijn LW, et al. Development of partial-thickness articular cartilage injury in a rabbit model. Clin Orthop Relat Res, 2008, 466(2) :487-494.
23. 王凡, 李爱冬, 羊惠君, 等. 转化生长因子-β对骺软骨细胞增殖和PCNA表达的影响. 华西医大学报, 2000, 31(4) :460-462, 467.
24. 任戈亮, 桂鉴超, 王黎明, 等. Ⅱ型胶原羧基端端肽对骨关节炎早期诊断和病情评估的实验研究. 南京医科大学学报(自然科学版), 2010, 30(5) :653-657.

Chinese Journal of Reparative and Reconstructive Surgery

MODEL ESTABLISHING OF PARTIAL-THICKNESS ARTICULAR CARTILAGE INJURY AND RELATIONSHIPS BETWEEN ACTIVATION OF CELLS AND EXPRESSION OF INTEGRIN β₁ IN A RAT MODEL

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Chinese Journal of Reparative and Reconstructive Surgery

MODEL ESTABLISHING OF PARTIAL-THICKNESS ARTICULAR CARTILAGE INJURY AND RELATIONSHIPS BETWEEN ACTIVATION OF CELLS AND EXPRESSION OF INTEGRIN β1 IN A RAT MODEL

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MODEL ESTABLISHING OF PARTIAL-THICKNESS ARTICULAR CARTILAGE INJURY AND RELATIONSHIPS BETWEEN ACTIVATION OF CELLS AND EXPRESSION OF INTEGRIN β₁ IN A RAT MODEL