Seeding fresh bone marrow aspirate directly on scaffolds to construct tissue-engineered ligament

doi:10.3969/j.issn.2095-4344.2014.12.011

Abstract

Abstract:

BACKGROUND: There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned.
OBJECTIVE: To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament
METHODS: We constructed fibroin fiber/small intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cells (bone marrow mesenchymal stem cells) on the scaffold to built cell seeding group. The control group had no treatment. After that, we detected the density of cell adhesion, cell proliferation ability and extracellular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks.
RESULTS AND CONCLUSION: After 4 hours of incubation, bone marrow seeding group was significantly higher than the cell seeding group in cell adhesion density and proliferation rate (P < 0.05). Bone marrow seeding group and cell seeding group showed higher type I, III collagen secretion compared with the control group (P < 0.05), but the collagen secretion of bone marrow seeding group and cell seeding group showed no significant difference. Composite cell scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: biocompatible materials, stents, ligaments, bone marrow

CLC Number:

R318

Wang Hong, Tang Ming, Meng Chun-qing, Wang Han-qi, Yang Shu-hua, Du Jing-yuan, Duan De-yu, Shao Zeng-wu. Seeding fresh bone marrow aspirate directly on scaffolds to construct tissue-engineered ligament[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(12): 1870-1876.

Figures/Tables 3

References

[1] Voigt C,Schönaich M,Lill H.Anterior cruciate ligament reconstruction: state of the art.Eur J Trauma.2006;32(4): 332-339.

[2] 敖英芳,田得祥,崔国庆,等.运动员前交叉韧带损伤的流行病学研究[J].体育科学,2000,20(4):47-49.

[3] Barenius B,Nordlander M,Ponzer S,et al. Quality of life and clinical outcome after anterior cruciate ligament reconstruction using patellar tendon graft or quadrupled semitendinosus graft: an 8-year follow-up of a randomized controlled trial.Am J Sports Med.2010;38(8):1533-1541.

[4] Longo UG,Buchmann S,Franceschetti E,et al.A systematic review of single-bundle versus double-bundle anterior cruciate ligament reconstruction.Br Med Bull. 2012;103(1): 147-168.

[5] Maffulli N,Spiezia F,King JB,et al. Figure-of-four pivot shift test: a technical note. Bull NYU Hosp Jt Dis.2011;69(2):173-176.

[6] Ge Z,Goh JCH,Lee EH.Selection of cell source for ligament tissue engineering.Cell Transplant.2005;14(8):573-583.

[7] Hoffmann A,Gross G.Tendon and ligament engineering in the adult organism: mesenchymal stem cells and gene-therapeutic approaches.Int Orthop. 2007;31(6): 791-797.

[8] Broese M,Toma I,Haasper C,et al.Seeding a human tendon matrix with bone marrow aspirates compared to previously isolated hBMSCs–An in vitro study.Technol Health Care.2011; 19(6):469-479.

[9] Abraham GA,Murray J,Billiar K,et al.Evaluation of the porcine intestinal collagen layer as a biomaterial.J Biomed Mater Res. 2000;51(3):442-452.

[10] 王成学,谷贵山,胡春明,等.老年骨质疏松动物模型韧带胶原蛋白含量测定[J].中国老年学杂志,2009,29(11):38-39.

[11] Grant RA.Estimation of hydroxyproline by the AutoAnalyser.J Clin Pathol.1964;17(6):685.

[12] Hankemeier S,Keus M,Zeichen J,et al.Modulation of proliferation and differentiation of human bone marrow stromal cells by fibroblast growth factor 2: potential implications for tissue engineering of tendons and ligaments. Tissue Eng.2005;11(1-2):41-49.

[13] Haasper C,Zeichen J,Meister R,et al.Tissue engineering of osteochondral constructs in vitro using bioreactors.Injury. 2008;39(1):66-76.

[14] Hernigou P,Daltro G,Filippini P,et al.Percutaneous implantation of autologous bone marrow osteoprogenitor cells as treatment of bone avascular necrosis related to sickle cell disease.Open Orthop J.2008;2:62.

[15] Asahara T,Masuda H,Takahashi T,et al.Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization.Circ Res.1999;85(3):221-228.

[16] Cristino S,Grassi F,Toneguzzi S,et al.Analysis of mesenchymal stem cells grown on a three‐dimensional HYAFF 11®‐based prototype ligament scaffold. J Biomed Mater Res A.2005;73(3):275-283.

[17] Garlet TP,Coelho U,Silva JS,et al.Cytokine expression pattern in compression and tension sides of the periodontal ligament during orthodontic tooth movement in humans.Eur J Oral Sci. 2007;115(5):355-362.

[18] 王洪,赵玉鑫,杨述华,等.半月板纤维软骨细胞与小肠粘膜下层相容性的研究[J].中华实验外科杂志,2007,24(9):1149-1149.

[19] 王茂源,赵建宁.小肠粘膜下层在骨科组织工程学中的进展[J].中国骨伤,2006,19(1):61-63.

[20] Liang R,Woo SLY,Takakura Y,et al.Long-term effects of porcine small intestine submucosa on the healing of medial collateral ligament: a functional tissue engineering study.J Orthop Res.2006;24(4):811-819.

[21] 王洪,李艳军,杨述华,等.单螺旋和3股螺旋蚕丝纤维支架的力学性能及其细胞黏附性能比较[J].中国组织工程研究与临床康复, 2007,11(18):3523-3526.

[22] 崔树北,王洪,杨述华,等.小肠黏膜下层包绕丝素纤维复合韧带的生物力学测定[J]. 中国组织工程研究与临床康复,2009,13(3): 517-520.

[23] Bertone AL,Goin S,Kamei SJ,et al.Metacarpophalangeal collateral ligament reconstruction using small intestinal submucosa in an equine model.J Biomed Mater Res A. 2008;84(1):219-229.