Growth Factor-conjugated Collagen Patch Prolongs Survival Rate of Transplanted Cells after Ventricular AneurysmRepair in Rats_Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Authors：

KANG Kai , QU Hui , TANG Jiquan , XIE Baodong , JIA Zhibo , ZHANG Yunan , WU Hua , LIRenke , JIANG Shulin.

1. Department of Cardiovascular Surgery，Second Affiliated Hospital of Harbin Medical University，Key Laboratory of Myocardial Ischemia Mechanism and Treatment established by Ministry of Education &;
Heilongjiang Province，Harbin 150086，P.R.China;;
2. Department of Pediatrics，Second Affiliated Hospital of Harbin Medical University，Harbin 150086，P. R. China;;
3. Department of Cardiothoracic Surgery，Jixi People’s Hospital， Jixi 158100，P. R. China;

Corresponding author：

Keywords：

Vascular endothelial growth factor; Basic fibroblast growth factor; Tissue engineering; Ventricular plasty

DOI：

10.7507/1007-4848.20130136

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Objective To observe the impact of collagen patches using 1-ethyl-3- （3-dimethylaminopropyl) carbod-iimide hydrochloride chemistry （EDC） to conjugate vascular endothelial growth factor （VEGF）＋ basic fibroblast growth factor （bFGF） or VEGF alone on the survival rate of transplanted human bone morrow mesenchymal stem cells （hBM-MSCs）in vitro and in vivo. Methods Collagen patches which were activated by EDC were used as the control group，and EDC activated collagen patches that were conjugated with VEGF or VEGF ＋ bFGF were used as the experiment groups（VEGF group and VEGF ＋ bFGF group）. hBM-MSCs （0.5×106/patch） were used as seeding cells to construct engineered heart tissue （EHT）. MTT assay was performed to assess in vitro proliferation of hBM-MSCs on 3 different collagen patches. Ventricular aneurysm model after myocardial infarction was created by left anterior descending artery （LAD） ligation in male SD rats，and EHT which were constructed with 3 different patches were used for ventricular plasty. Four weeks later，immunofluorescence staining was used to examine arteriole density （anti-α-SMA staining） and transplanted cell survival （anti-h-mitochondria staining）. Results （1） hMSCs proliferation in VEGF group and VEGF ＋ bFGF group was significantly better than that in the control group on the 2nd and 4th day after cell transplantation （P＜0.05）；（2） Four weeks afterEHT implantation，immunofluorescence staining for α-SMA revealed that arteriole density of VEGF group and VEGF ＋ bFGF group was significantly higher than that of the control group （P＜0.05）；（3） Immunofluorescence staining forh-mitochondria showed that survival rates of transplanted hBM-MSCs of VEGF group and VEGF ＋ bFGF group were significantly higher than that of the control group （P＜0.05）；（4） There was a significantly positive correlation between survival rate of hBM-MSCs and arteriole density （r 2=0.99，P=0.02）. Conclusion VEGF or VEGF ＋ bFGF conjugated collagen patch can significantly improve hBM-MSCs proliferation in vitro and enhance survival rate of transplanted hBM-MSCs by accelerating revascularization of EHT in vivo.

Citation： KANG Kai,QU Hui,TANG Jiquan,XIE Baodong,JIA Zhibo,ZHANG Yunan,WU Hua,LIRenke,JIANG Shulin.. Growth Factor-conjugated Collagen Patch Prolongs Survival Rate of Transplanted Cells after Ventricular AneurysmRepair in Rats. Chinese Journal of Clinical Thoracic and Cardiovascular Surgery, 2013, 20(4): 451-456. doi: 10.7507/1007-4848.20130136 Copy

1.	Ozawa T， Mickle DA， Weisel RD， et al. Histologic changes of nonbiodegradable and biodegradable biomaterials used to repair right ventricular heart defects in rats. J Thorac Cardiovasc Surg， 2002， 124 （6）：1157-1164.
2.	Miyagi Y， Zeng F， Huang XP， et al. Surgical ventricular restoration with a cell- and cytokine-seeded biodegradable scaffold. Biomaterials，2010， 31 （30）：7684-7694.
3.	Naderi H， Matin MM， Bahrami AR. Review paper：critical issues in tissue engineering：biomaterials， cell sources， angiogenesis， and drug delivery systems. J Biomater Appl， 2011， 26 （4）：383-417.
4.	Simpson D， Liu H， Fan TH， et al. A tissue engineering approach to progenitor cell delivery results in significant cell engraftment and improved myocardial remodeling. Stem Cells， 2007， 25 （9）：2350-2357.
5.	康凯，曲辉，汤继权，等. 大鼠室壁瘤左心室重建模型的建立. 中国胸心血管外科临床杂志， 2012， 19 （3）：293-297.
6.	Radisic M， Malda J， Epping E， et al. Oxygen gradients correlate with cell density and cell viability in engineered cardiac tissue.Biotechnol Bioeng， 2006， 93 （2）：332-343.
7.	Miyagi Y， Chiu LL， Cimini M， et al. Biodegradable collagen patch with covalently immobilized VEGF for myocardial repair. Biomaterials，2011， 32 （5）：1280-1290.
8.	Wang H， Zhang X， Li Y， et al. Improved myocardial performance in infarcted rat heart by co-injection of basic fibroblast growth factor with temperature-responsive chitosan hydrogel. J Heart Lung Transplant， 2010， 29 （8）：881-887.
9.	Sun Q， Silva EA， Wang A， et al. Sustained release of multiple growth factors from injectable polymeric system as a novel therapeuticapproach towards angiogenesis. Pharm Res， 2010， 27 （2）：264-271.
10.	Lee KY， Peters MC， Anderson KW， et al. Controlled growth factorrelease from synthetic extracellular matrices. Nature， 2000， 408 （6815）：998-1000.
11.	Chiu LL， Radisic M. Scaffolds with covalently immobilized VEGF and Angiopoietin-1 for vascularization of engineered tissues. Biomaterials， 2010， 31 （2）：226-241.
12.	Zheng W， Seftor EA， Meininger CJ， et al. Mechanisms of coronary angiogenesis in response to stretch：role of VEGF and TGF-beta. Am J Physiol Heart Circ Physiol， 2001， 280 （2）：H909-H917.
13.	Oh H， Bradfute SB， Gallardo TD， et al. Cardiac progenitor cells from adult myocardium：homing， differentiation， and fusion afterinfarction. Proc Natl Acad Sci U S A， 2003， 100 （21）：12313-12318.
14.	Asahara T， Bauters C， Zheng LP， et al. Synergistic effect of vascularendothelial growth factor and basic fibroblast growth factor on angiogenesis in vivo. Circulation， 1995， 92 （9 Suppl）：Ⅱ 365- Ⅱ 371.
15.	Onimaru M， Yonemitsu Y， Tanii M， et al. Fibroblast growth factor-2gene transfer can stimulate hepatocyte growth factor expression irrespective of hypoxia-mediated downregulation in ischemic limbs. Circ Res， 2002， 91 （10）：923-930.
16.	Lee JS， Kim JM， Kim KL， et al. Combined administration of naked DNA vectors encoding VEGF and bFGF enhances tissue perfusion and arteriogenesis in ischemic hindlimb. Biochem Biophys Res Commun，2007， 360 （4）：752-758.
17.	Spanholtz TA， Theodorou P， Holzbach T， et al. Vascular endothelialgrowth factor （VEGF165） plus basic fibroblast growth factor （bFGF） producing cells induce a mature and stable vascular network—afuture therapy for ischemically challenged tissue. J Surg Res， 2011， 171 （1）：329-338.
18.	Dvir T， Kedem A， Ruvinov E， et al. Prevascularization of cardiac patch on the omentum improves its therapeutic outcome. Proc Natl Acad Sci U S A， 2009， 106 （35）：14990-14995.
19.	Zhuo Y， Li SH， Chen MS， et al. Aging impairs the angiogenic response to ischemic injury and the activity of implanted cells：combined Consequences for cell therapy in older recipients. J Thorac Cardiovasc Surg， 2010， 139 （5）：1286-1294.

1. 　Ozawa T， Mickle DA， Weisel RD， et al. Histologic changes of nonbiodegradable and biodegradable biomaterials used to repair right ventricular heart defects in rats. J Thorac Cardiovasc Surg， 2002， 124 （6）：1157-1164.
2. 　Miyagi Y， Zeng F， Huang XP， et al. Surgical ventricular restoration with a cell- and cytokine-seeded biodegradable scaffold. Biomaterials，2010， 31 （30）：7684-7694.
3. 　Naderi H， Matin MM， Bahrami AR. Review paper：critical issues in tissue engineering：biomaterials， cell sources， angiogenesis， and drug delivery systems. J Biomater Appl， 2011， 26 （4）：383-417.
4. 　Simpson D， Liu H， Fan TH， et al. A tissue engineering approach to progenitor cell delivery results in significant cell engraftment and improved myocardial remodeling. Stem Cells， 2007， 25 （9）：2350-2357.
5. 　康凯，曲辉，汤继权，等. 大鼠室壁瘤左心室重建模型的建立. 中国胸心血管外科临床杂志， 2012， 19 （3）：293-297.
6. 　Radisic M， Malda J， Epping E， et al. Oxygen gradients correlate with cell density and cell viability in engineered cardiac tissue.Biotechnol Bioeng， 2006， 93 （2）：332-343.
7. 　Miyagi Y， Chiu LL， Cimini M， et al. Biodegradable collagen patch with covalently immobilized VEGF for myocardial repair. Biomaterials，2011， 32 （5）：1280-1290.
8. 　Wang H， Zhang X， Li Y， et al. Improved myocardial performance in infarcted rat heart by co-injection of basic fibroblast growth factor with temperature-responsive chitosan hydrogel. J Heart Lung Transplant， 2010， 29 （8）：881-887.
9. 　Sun Q， Silva EA， Wang A， et al. Sustained release of multiple growth factors from injectable polymeric system as a novel therapeuticapproach towards angiogenesis. Pharm Res， 2010， 27 （2）：264-271.
10. 　Lee KY， Peters MC， Anderson KW， et al. Controlled growth factorrelease from synthetic extracellular matrices. Nature， 2000， 408 （6815）：998-1000.
11. 　Chiu LL， Radisic M. Scaffolds with covalently immobilized VEGF and Angiopoietin-1 for vascularization of engineered tissues. Biomaterials， 2010， 31 （2）：226-241.
12. 　Zheng W， Seftor EA， Meininger CJ， et al. Mechanisms of coronary angiogenesis in response to stretch：role of VEGF and TGF-beta. Am J Physiol Heart Circ Physiol， 2001， 280 （2）：H909-H917.
13. 　Oh H， Bradfute SB， Gallardo TD， et al. Cardiac progenitor cells from adult myocardium：homing， differentiation， and fusion afterinfarction. Proc Natl Acad Sci U S A， 2003， 100 （21）：12313-12318.
14. 　Asahara T， Bauters C， Zheng LP， et al. Synergistic effect of vascularendothelial growth factor and basic fibroblast growth factor on angiogenesis in vivo. Circulation， 1995， 92 （9 Suppl）：Ⅱ 365- Ⅱ 371.
15. 　Onimaru M， Yonemitsu Y， Tanii M， et al. Fibroblast growth factor-2gene transfer can stimulate hepatocyte growth factor expression irrespective of hypoxia-mediated downregulation in ischemic limbs. Circ Res， 2002， 91 （10）：923-930.
16. 　Lee JS， Kim JM， Kim KL， et al. Combined administration of naked DNA vectors encoding VEGF and bFGF enhances tissue perfusion and arteriogenesis in ischemic hindlimb. Biochem Biophys Res Commun，2007， 360 （4）：752-758.
17. 　Spanholtz TA， Theodorou P， Holzbach T， et al. Vascular endothelialgrowth factor （VEGF165） plus basic fibroblast growth factor （bFGF） producing cells induce a mature and stable vascular network—afuture therapy for ischemically challenged tissue. J Surg Res， 2011， 171 （1）：329-338.
18. 　Dvir T， Kedem A， Ruvinov E， et al. Prevascularization of cardiac patch on the omentum improves its therapeutic outcome. Proc Natl Acad Sci U S A， 2009， 106 （35）：14990-14995.
19. 　Zhuo Y， Li SH， Chen MS， et al. Aging impairs the angiogenic response to ischemic injury and the activity of implanted cells：combined Consequences for cell therapy in older recipients. J Thorac Cardiovasc Surg， 2010， 139 （5）：1286-1294.

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Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

Growth Factor-conjugated Collagen Patch Prolongs Survival Rate of Transplanted Cells after Ventricular AneurysmRepair in Rats

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