EFFECT OF HAMSTRING TENDON TRANSFECTED WITH ADENOVIRUSMEDIATED TRANSFORMING GROWTH FACTOR β<sub>1</sub> GENE ON HISTOMORPHOLOGY OF TENDON-BONE INTERFACE HEALING AFTER ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION IN RABBITS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

 WANGXiaoxu , ZHAIRongfan , YANGJuntao , ZHOUShouhong , TANWenfu , TANGuanghua

Department of Orthopaedics, the Second Affiliated Hospital of Nanhua University, Hengyang Hunan, 421001, P. R. China;

Corresponding author：

WANGXiaoxu, Email: wxx1024@163.com

Keywords：

Adenovirus; Transforming growth factor β₁; Anterior cruciate ligament; Tendon-bone healing; Rabbit

DOI：

10.7507/1002-1892.20150319

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Objective To investigate the effect of hamstring tendon transfected with adenovirus-mediated transforming growth factor β₁ (AdTGF-β₁) genes on the histomorphology of tendon-bone interface healing after anterior cruciate ligament (ACL) reconstruction in rabbits. Methods AdTGF-β₁ and AdGFP were diluted to 5×10⁸ PFU/mL with DMEM. Forty-eight New Zealand white rabbits were divided into 3 groups randomly (n=16), weighing 1.6-2.5 kg for ACL reconstruction with hamstring tendon autograft. Hamstring tendon was cultured and transfected with AdTGF-β₁ (group A) and AdGFP (group B) for 12 hours before ACL reconstruction, and was cultured with DMEM in group C. After 12 hours of transfection, the expression of green fluorescence was observed in groups A and B under fluorescence microscopy; TGF-β₁ protein level was detected by ELISA in group A. At 2, 4, 8, and 12 weeks after operation, the specimens were harvested for HE and Masson staining; the number of fibroblasts was counted, and the Buark grading was used to evaluate tendon-bone interface healing. Results Green fluorescence was observed after 12 hours of transfection in groups A and B. TGF-β₁ protein level reached (221.0±12.2) ng/mL at 12 hours in group A. The histological observation showed that few fibroblasts and collagen fibers were found, and Sharpey fibers appeared in group A; regular Sharpey fibers were seen in the interface, and integrity interface in some areas at 12 weeks. But fibroblasts of groups B and C were less than those of group A, with loose tendon-bone interface; no integrity interface was observed at 12 weeks. The number of fibroblasts and Buark grading of group A were significantly higher than those of groups B and C (P<0.05), but no significant difference was found between groups B and C (P>0.05). Conclusion Hamstring tendon transfected with AdTGF-β₁ gene can promote the healing of tendon-bone interface after ACL reconstruction.

Citation： WANGXiaoxu, ZHAIRongfan, YANGJuntao, ZHOUShouhong, TANWenfu, TANGuanghua. EFFECT OF HAMSTRING TENDON TRANSFECTED WITH ADENOVIRUSMEDIATED TRANSFORMING GROWTH FACTOR β₁ GENE ON HISTOMORPHOLOGY OF TENDON-BONE INTERFACE HEALING AFTER ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION IN RABBITS. Chinese Journal of Reparative and Reconstructive Surgery, 2015, 29(12): 1488-1493. doi: 10.7507/1002-1892.20150319 Copy

1.	Arnoczky SP, Tarvin GB, Marshall JL. Anterior cruciate ligament replacement using patellar tendon. An evaluation of graft revascularization in the dog. J Bone Joint Surg (Am), 1982, 64(2):217-224.
2.	Scranton PE Jr, Lanzer WL, Ferguson MS, et al. Mechanisms of anterior cruciate ligament neovascularization and ligamentization. Arthroscopy, 1998, 14(7):702-716.
3.	Murray MM, Fleming BC. Use of a bioactive scaffold to stimulate anterior cruciate ligament healing also minimizes posttraumatic osteoarthritis after surgery. Am J Sports Med, 2013, 41(8):1762-1770.
4.	Uozumi H, Aizawa T, Sugita T, et al. Visualization of torn anterior cruciate ligament using 3-dimensional computed tomography. Orthop Rev (Pavia), 2013, 5(3):e22.
5.	Bushnell BD, Sakryd G, Noonan TJ. Hamstring donor-site block:evaluation of pain control after anterior cruciate ligament reconstruction. Arthroscopy, 2010, 26(7):894-900.
6.	Nguyen DT, Ramwadhdoebe TH, van der Hart CP, et al. Intrinsic healing response of the human anterior cruciate ligament:an histological study of teattached ACL rremnants. J Orthop Res, 2014, 32(2):296-301.
7.	Rodeo SA, Voigt C, Ma R,et al. Use of a new model allowing controlled uniaxial loading to evaluate tendon healing in a bone tunnel. J Orthop Res, 2015, 10(2):1-29.
8.	Riboh JC, Hasselblad V, Godin JA, et al. Transtibial versus independent drilling techniques for anterior cruciate ligament reconstruction:a systematic review, meta-analysis, and metaregression. Am J Sports Med, 2013, 41(11):2693-2702.
9.	Lefevre N, Naouri JF, Bohu Y, et al. Partial tears of the anterior cruciate ligament:diagnostic performance of isotropic threedimensional fast spin echo (3D-FSE-Cube) MRI. Eur J Orthop Surg Traumatol, 2014, 24(1):85-91.
10.	Wei X, Mao Z, Hou Y, et al. Local administration of TGFβ-1/VEGF165 gene-transduced bone mesenchymal stem cells for Achilles allograft replacement of the anterior cruciate ligament in rabbits. Biochem Biophys Res Commun, 2011, 406(2):204-210.
11.	谢磊, 陈百成, 王晓峰, 等. 自体及同种异体肌腱重建前交叉韧带术后本体感觉恢复的比较研究. 中国修复重建外科杂志, 2011, 25(8):907-911.
12.	Colvin A, Sharma C, Parides M, et al. What is the best femoral fixation of hamstring autografts in anterior cruciate ligament reconstruction?:a meta-analysis. Clin Orthop Relat Res, 2011, 469(4):1075-1081.
13.	Lee GH, McCulloch P, Cole BJ, et al. The incidence of acute patellar tendon harvest complications for anterior cruciate ligament reconstruction. Arthroscopy, 2008, 24(2):262-166.
14.	Tan J, Hu Y, Zheng H, et al. Construction of recombinant adenoviral vector Ad-CMV-hTGFbeta1 for reversion of intervertebral disc degeneration by gene transfer. Chin J Traumatol, 2002, 5(2):97-102.
15.	Gao X, Du J, Fan G, et al. Adenoviral-mediated GDNF protects bone marrow mesenchymal stem cells against apoptosis induced by hydrogen peroxide. Biomed Mater Eng, 2014, 24(6):2169-2176.
16.	Bissell L, Tibrewal S, Sahn V, et al. Growth factors and platelet rich plasma in anterior cruciate ligament reconstruction. Curr Stem Cell Res Ther, 2014, 10(1):19-25.
17.	Marcacci M, Marcheggiani Muccioli GM, Grassi A, et al. Arthroscopic meniscus allograft transplantation in male professional soccer players:a 36-month follow-up study. Am J Sports Med, 2014, 42(2):382-388.

1. Arnoczky SP, Tarvin GB, Marshall JL. Anterior cruciate ligament replacement using patellar tendon. An evaluation of graft revascularization in the dog. J Bone Joint Surg (Am), 1982, 64(2):217-224.
2. Scranton PE Jr, Lanzer WL, Ferguson MS, et al. Mechanisms of anterior cruciate ligament neovascularization and ligamentization. Arthroscopy, 1998, 14(7):702-716.
3. Murray MM, Fleming BC. Use of a bioactive scaffold to stimulate anterior cruciate ligament healing also minimizes posttraumatic osteoarthritis after surgery. Am J Sports Med, 2013, 41(8):1762-1770.
4. Uozumi H, Aizawa T, Sugita T, et al. Visualization of torn anterior cruciate ligament using 3-dimensional computed tomography. Orthop Rev (Pavia), 2013, 5(3):e22.
5. Bushnell BD, Sakryd G, Noonan TJ. Hamstring donor-site block:evaluation of pain control after anterior cruciate ligament reconstruction. Arthroscopy, 2010, 26(7):894-900.
6. Nguyen DT, Ramwadhdoebe TH, van der Hart CP, et al. Intrinsic healing response of the human anterior cruciate ligament:an histological study of teattached ACL rremnants. J Orthop Res, 2014, 32(2):296-301.
7. Rodeo SA, Voigt C, Ma R,et al. Use of a new model allowing controlled uniaxial loading to evaluate tendon healing in a bone tunnel. J Orthop Res, 2015, 10(2):1-29.
8. Riboh JC, Hasselblad V, Godin JA, et al. Transtibial versus independent drilling techniques for anterior cruciate ligament reconstruction:a systematic review, meta-analysis, and metaregression. Am J Sports Med, 2013, 41(11):2693-2702.
9. Lefevre N, Naouri JF, Bohu Y, et al. Partial tears of the anterior cruciate ligament:diagnostic performance of isotropic threedimensional fast spin echo (3D-FSE-Cube) MRI. Eur J Orthop Surg Traumatol, 2014, 24(1):85-91.
10. Wei X, Mao Z, Hou Y, et al. Local administration of TGFβ-1/VEGF165 gene-transduced bone mesenchymal stem cells for Achilles allograft replacement of the anterior cruciate ligament in rabbits. Biochem Biophys Res Commun, 2011, 406(2):204-210.
11. 谢磊, 陈百成, 王晓峰, 等. 自体及同种异体肌腱重建前交叉韧带术后本体感觉恢复的比较研究. 中国修复重建外科杂志, 2011, 25(8):907-911.
12. Colvin A, Sharma C, Parides M, et al. What is the best femoral fixation of hamstring autografts in anterior cruciate ligament reconstruction?:a meta-analysis. Clin Orthop Relat Res, 2011, 469(4):1075-1081.
13. Lee GH, McCulloch P, Cole BJ, et al. The incidence of acute patellar tendon harvest complications for anterior cruciate ligament reconstruction. Arthroscopy, 2008, 24(2):262-166.
14. Tan J, Hu Y, Zheng H, et al. Construction of recombinant adenoviral vector Ad-CMV-hTGFbeta1 for reversion of intervertebral disc degeneration by gene transfer. Chin J Traumatol, 2002, 5(2):97-102.
15. Gao X, Du J, Fan G, et al. Adenoviral-mediated GDNF protects bone marrow mesenchymal stem cells against apoptosis induced by hydrogen peroxide. Biomed Mater Eng, 2014, 24(6):2169-2176.
16. Bissell L, Tibrewal S, Sahn V, et al. Growth factors and platelet rich plasma in anterior cruciate ligament reconstruction. Curr Stem Cell Res Ther, 2014, 10(1):19-25.
17. Marcacci M, Marcheggiani Muccioli GM, Grassi A, et al. Arthroscopic meniscus allograft transplantation in male professional soccer players:a 36-month follow-up study. Am J Sports Med, 2014, 42(2):382-388.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF HAMSTRING TENDON TRANSFECTED WITH ADENOVIRUSMEDIATED TRANSFORMING GROWTH FACTOR β₁ GENE ON HISTOMORPHOLOGY OF TENDON-BONE INTERFACE HEALING AFTER ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION IN RABBITS

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EFFECT OF HAMSTRING TENDON TRANSFECTED WITH ADENOVIRUSMEDIATED TRANSFORMING GROWTH FACTOR β1 GENE ON HISTOMORPHOLOGY OF TENDON-BONE INTERFACE HEALING AFTER ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION IN RABBITS

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EFFECT OF HAMSTRING TENDON TRANSFECTED WITH ADENOVIRUSMEDIATED TRANSFORMING GROWTH FACTOR β₁ GENE ON HISTOMORPHOLOGY OF TENDON-BONE INTERFACE HEALING AFTER ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION IN RABBITS