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,劉超, 等. 早期兔骨關節炎軟骨CD105+/CD166+細胞及其軟骨分化潛能的實驗研究. 中國修復重建外科雜志, 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, Kirst?tter E, et al. β₁ integrins mediate attachment of mesenchymal stem cells to cartilage lesions. Biores Open Access, 2015, 4(1) :39-53.
17.	閆德雄, 蘇踴躍, 唐紅英, 等. 整合素β₁在人表皮細胞株HaCaT增殖中影響的研究. 第三軍醫大學學報, 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.	張向鑫, 馬瑞雪, 李天友, 等. 整合素β₁在髖關節發育不良髖臼軟骨退變中的作用. 臨床小兒外科雜志, 2009, 8(3) :29-33.
21.	鐘清玲, 劉德伍, 劉繁榮, 等. 糖尿病大鼠表皮干細胞及其增殖分化相關蛋白的研究. 中國修復重建外科雜志, 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, 李愛冬, 羊惠君, 等. 轉化生長因子-β對骺軟骨細胞增殖和PCNA表達的影響. 華西醫大學報, 2000, 31(4) :460-462, 467.
24.	任戈亮, 桂鑒超, 王黎明, 等. Ⅱ型膠原羧基端端肽對骨關節炎早期診斷和病情評估的實驗研究. 南京醫科大學學報(自然科學版), 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,劉超, 等. 早期兔骨關節炎軟骨CD105+/CD166+細胞及其軟骨分化潛能的實驗研究. 中國修復重建外科雜志, 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, Kirst?tter E, et al. β₁ integrins mediate attachment of mesenchymal stem cells to cartilage lesions. Biores Open Access, 2015, 4(1) :39-53.
17. 閆德雄, 蘇踴躍, 唐紅英, 等. 整合素β₁在人表皮細胞株HaCaT增殖中影響的研究. 第三軍醫大學學報, 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. 張向鑫, 馬瑞雪, 李天友, 等. 整合素β₁在髖關節發育不良髖臼軟骨退變中的作用. 臨床小兒外科雜志, 2009, 8(3) :29-33.
21. 鐘清玲, 劉德伍, 劉繁榮, 等. 糖尿病大鼠表皮干細胞及其增殖分化相關蛋白的研究. 中國修復重建外科雜志, 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, 李愛冬, 羊惠君, 等. 轉化生長因子-β對骺軟骨細胞增殖和PCNA表達的影響. 華西醫大學報, 2000, 31(4) :460-462, 467.
24. 任戈亮, 桂鑒超, 王黎明, 等. Ⅱ型膠原羧基端端肽對骨關節炎早期診斷和病情評估的實驗研究. 南京醫科大學學報(自然科學版), 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