Study on injectable chitosan hydrogel with tendon-derived stem cells for enhancing rotator cuff tendon-to-bone healing_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

WEN Huawei ^1,2 , ZHANG Qingsong ^1,2 , TANG Ming ^1,2 , LI Ya’nan ^1,2 , TAN Hongfei ^1,2 ,  FANG Yushun ^1,2

1. Department of Sport Medicine, Wuhan Fourth Hospital, Wuhan Hubei, 430030, P. R. China;
2. Hubei Sports Medicine Center, Wuhan Hubei, 430030, P. R. China;

Corresponding author：

FANG Yushun, Email: 007fq@163.com

Keywords：

Rotator cuff repair; tendon-derived stem cells; chitosan; hydrogel; tendon-to-bone healing; rabbit

DOI：

10.7507/1002-1892.202309014

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To explore the effect of chitosan (CS) hydrogel loaded with tendon-derived stem cells (TDSCs; hereinafter referred to as TDSCs/CS hydrogel) on tendon-to-bone healing after rotator cuff repair in rabbits. Methods TDSCs were isolated from the rotator cuff tissue of 3 adult New Zealand white rabbits by Henderson step-by-step enzymatic digestion method and identified by multidirectional differentiation and flow cytometry. The 3rd generation TDSCs were encapsulated in CS to construct TDSCs/CS hydrogel. The cell counting kit 8 (CCK-8) assay was used to detect the proliferation of TDSCs in the hydrogel after 1-5 days of culture in vitro, and cell compatibility of TDSCs/CS hydrogel was evaluated by using TDSCs alone as control. Another 36 adult New Zealand white rabbits were randomly divided into 3 groups (n=12): rotator cuff repair group (control group), rotator cuff repair+CS hydrogel injection group (CS group), and rotator cuff repair+TDSCs/CS hydrogel injection group (TDSCs/CS group). After establishing the rotator cuff repair models, the corresponding hydrogel was injected into the tendon-to-bone interface in the CS group and TDSCs/CS group, and no other treatment was performed in the control group. The general condition of the animals was observed after operation. At 4 and 8 weeks, real-time quantitative PCR (qPCR) was used to detect the relative expressions of tendon forming related genes (tenomodulin, scleraxis), chondrogenesis related genes (aggrecan, sex determining region Y-related high mobility group-box gene 9), and osteogenesis related genes (alkaline phosphatase, Runt-related transcription factor 2) at the tendon-to-bone interface. At 8 weeks, HE and Masson staining were used to observe the histological changes, and the biomechanical test was used to evaluate the ultimate load and the failure site of the repaired rotator cuff to evaluate the tendon-to-bone healing and biomechanical properties. Results CCK-8 assay showed that the CS hydrogel could promote the proliferation of TDSCs (P<0.05). qPCR results showed that the expressions of tendon-to-bone interface related genes were significantly higher in the TDSCs/CS group than in the CS group and control group at 4 and 8 weeks after operation (P<0.05). Moreover, the expressions of tendon-to-bone interface related genes at 8 weeks after operation were significantly higher than those at 4 weeks after operation in the TDSCs/CS group (P<0.05). Histological staining showed the clear cartilage tissue and dense and orderly collagen formation at the tendon-to-bone interface in the TDSCs/CS group. The results of semi-quantitative analysis showed that compared with the control group, the number of cells, the proportion of collagen fiber orientation, and the histological score in the TDSCs/CS group increased, the vascularity decreased, showing significant differences (P<0.05); compared with the CS group, the proportion of collagen fiber orientation and the histological score in the TDSCs/CS group significantly increased (P<0.05), while there was no significant difference in the number of cells and vascularity (P>0.05). All samples in biomechanical testing failed at the repair site during the testing process. The ultimate load of the TDSCs/CS group was significantly higher than that of the control group (P<0.05), but there was no significant difference compared to the CS group (P>0.05). Conclusion TDSCs/CS hydrogel can induce cartilage regeneration to promote rotator cuff tendon-to-bone healing.

Citation： WEN Huawei, ZHANG Qingsong, TANG Ming, LI Ya’nan, TAN Hongfei, FANG Yushun. Study on injectable chitosan hydrogel with tendon-derived stem cells for enhancing rotator cuff tendon-to-bone healing. Chinese Journal of Reparative and Reconstructive Surgery, 2024, 38(1): 91-98. doi: 10.7507/1002-1892.202309014 Copy

1.	Goutallier D, Postel JM, Gleyze P, et al. Influence of cuff muscle fatty degeneration on anatomic and functional outcomes after simple suture of full-thickness tears. J Shoulder Elbow Surg, 2003, 12(6): 550-554.
2.	Cai Z, Zhang Y, Liu S, et al. Celecoxib, beyond anti-inflammation, alleviates tendon-derived stem cell senescence in degenerative rotator cuff tendinopathy. Am J Sports Med, 2022, 50(9): 2488-2496.
3.	Plancher KD, Shanmugam J, Briggs K, et al. Diagnosis and management of partial thickness rotator cuff tears: a comprehensive review. J Am Acad Orthop Surg, 2021, 29(24): 1031-1043.
4.	汤明, 文华伟, 张绍华, 等. 阔筋膜补片桥接联合肱二头肌长头腱转位治疗不可修复性肩袖撕裂. 中华骨科杂志, 2023, 43(4): 238-246.
5.	Galetta MD, Keller RE, Sabbag OD, et al. Rehabilitation variability after rotator cuff repair. J Shoulder Elbow Surg, 2021, 30(6): e322-e333.
6.	Burkhart SS, Diaz Pagàn JL, Wirth MA, et al. Cyclic loading of anchor-based rotator cuff repairs: confirmation of the tension overload phenomenon and comparison of suture anchor fixation with transosseous fixation. Arthroscopy, 1997, 13(6): 720-724.
7.	Goradia VK, Mullen DJ, Boucher HR, et al. Cyclic loading of rotator cuff repairs: A comparison of bioabsorbable tacks with metal suture anchors and transosseous sutures. Arthroscopy, 2001, 17(4): 360-364.
8.	Kim SJ, Song DH, Park JW, et al. Effect of bone marrow aspirate concentrate-platelet-rich plasma on tendon-derived stem cells and rotator cuff tendon tear. Cell Transplant, 2017, 26(5): 867-878.
9.	Lui PP, Chan KM. Tendon-derived stem cells (TDSCs): from basic science to potential roles in tendon pathology and tissue engineering applications. Stem Cell Rev Rep, 2011, 7(4): 883-897.
10.	Yu HB, Xiong J, Zhang HZ, et al. TGFβ1-transfected tendon stem cells promote tendon fibrosis. J Orthop Surg Res, 2022, 17(1): 358.
11.	Cheng B, Ge H, Zhou J, et al. TSG-6 mediates the effect of tendon derived stem cells for rotator cuff healing. Eur Rev Med Pharmacol Sci, 2014, 18(2): 247-251.
12.	Shen H, Cheng L, Zheng Q, et al. Scavenging of reactive oxygen species can adjust the differentiation of tendon stem cells and progenitor cells and prevent ectopic calcification in tendinopathy. Acta Biomater, 2022, 152: 440-452.
13.	Zhao J, Qiu P, Wang Y, et al. Chitosan-based hydrogel wound dressing: From mechanism to applications, a review. Int J Biol Macromol, 2023, 244: 125250.
14.	Guadarrama-Escobar OR, Serrano-Castañeda P, Anguiano-Almazán E, et al. Chitosan nanoparticles as oral drug carriers. Int J Mol Sci, 2023, 24(5): 4289.
15.	Jafernik K, Ładniak A, Blicharska E, et al. Chitosan-based nanoparticles as effective drug delivery systems-A review. Molecules, 2023, 28(4): 1963.
16.	Li X, Su Z, Shen K, et al. Eugenol-preconditioned mesenchymal stem cell-derived extracellular vesicles promote antioxidant capacity of tendon stem cells in vitro and in vivo. Oxid Med Cell Longev, 2022, 2022: 3945195.
17.	Wang Y, Zhao Y, Ma S, et al. Injective programmable proanthocyanidin coordinated zinc-based composite hydrogel for infected bone repair. Adv Healthc Mater, 2023, 27: e202302690.
18.	Chen P, Cui L, Fu SC, et al. The 3D-printed PLGA scaffolds loaded with bone marrow-derived mesenchymal stem cells augment the healing of rotator cuff repair in the rabbits. Cell Transplant, 2020, 29: 963689720973647.
19.	Rothrauff BB, Smith CA, Ferrer GA, et al. The effect of adipose-derived stem cells on enthesis healing after repair of acute and chronic massive rotator cuff tears in rats. J Shoulder Elbow Surg, 2019, 28(4): 654-664.
20.	Cao Y, Yang S, Zhao D, et al. Three-dimensional printed multiphasic scaffolds with stratified cell-laden gelatin methacrylate hydrogels for biomimetic tendon-to-bone interface engineering. J Orthop Translat, 2020, 23: 89-100.
21.	Shin MJ, Shim IK, Kim DM, et al. Engineered cell sheets for the effective delivery of adipose-derived stem cells for tendon-to-bone healing. Am J Sports Med, 2020, 48(13): 3347-3358.
22.	Liu Q, Hatta T, Qi J, et al. Novel engineered tendon-fibrocartilage-bone composite with cyclic tension for rotator cuff repair. J Tissue Eng Regen Med, 2018, 12(7): 1690-1701.
23.	Rui YF, Lui PP, Lee YW, et al. Higher BMP receptor expression and BMP-2-induced osteogenic differentiation in tendon-derived stem cells compared with bone-marrow-derived mesenchymal stem cells. Int Orthop, 2012, 36(5): 1099-1107.
24.	Mandalia K, Mousad A, Welborn B, et al. Scaffold- and graft-based biological augmentation of rotator cuff repair: an updated systematic review and meta-analysis of preclinical and clinical studies for 2010-2022. J Shoulder Elbow Surg, 2023, 32(9): 1784-1800.
25.	Credille KT, Wang ZRC, Horner NS, et al. Biphasic interpositional allograft for rotator cuff repair augmentation is safe in an ovine model. Arthroscopy, 2023, 39(9): 1983-1997.
26.	Abourehab MAS, Pramanik S, Abdelgawad MA, et al. Recent advances of chitosan formulations in biomedical applications. Int J Mol Sci, 2022, 23(18): 10975.
27.	Willbold E, Wellmann M, Welke B, et al. Possibilities and limitations of electrospun chitosan-coated polycaprolactone grafts for rotator cuff tear repair. J Tissue Eng Regen Med, 2020, 14(1): 186-197.
28.	Niu Y, Wu J, Kang Y, et al. Recent advances of magnetic chitosan hydrogel: Preparation, properties and applications. Int J Biol Macromol, 2023, 247: 125722.

1. Goutallier D, Postel JM, Gleyze P, et al. Influence of cuff muscle fatty degeneration on anatomic and functional outcomes after simple suture of full-thickness tears. J Shoulder Elbow Surg, 2003, 12(6): 550-554.
2. Cai Z, Zhang Y, Liu S, et al. Celecoxib, beyond anti-inflammation, alleviates tendon-derived stem cell senescence in degenerative rotator cuff tendinopathy. Am J Sports Med, 2022, 50(9): 2488-2496.
3. Plancher KD, Shanmugam J, Briggs K, et al. Diagnosis and management of partial thickness rotator cuff tears: a comprehensive review. J Am Acad Orthop Surg, 2021, 29(24): 1031-1043.
4. 汤明, 文华伟, 张绍华, 等. 阔筋膜补片桥接联合肱二头肌长头腱转位治疗不可修复性肩袖撕裂. 中华骨科杂志, 2023, 43(4): 238-246.
5. Galetta MD, Keller RE, Sabbag OD, et al. Rehabilitation variability after rotator cuff repair. J Shoulder Elbow Surg, 2021, 30(6): e322-e333.
6. Burkhart SS, Diaz Pagàn JL, Wirth MA, et al. Cyclic loading of anchor-based rotator cuff repairs: confirmation of the tension overload phenomenon and comparison of suture anchor fixation with transosseous fixation. Arthroscopy, 1997, 13(6): 720-724.
7. Goradia VK, Mullen DJ, Boucher HR, et al. Cyclic loading of rotator cuff repairs: A comparison of bioabsorbable tacks with metal suture anchors and transosseous sutures. Arthroscopy, 2001, 17(4): 360-364.
8. Kim SJ, Song DH, Park JW, et al. Effect of bone marrow aspirate concentrate-platelet-rich plasma on tendon-derived stem cells and rotator cuff tendon tear. Cell Transplant, 2017, 26(5): 867-878.
9. Lui PP, Chan KM. Tendon-derived stem cells (TDSCs): from basic science to potential roles in tendon pathology and tissue engineering applications. Stem Cell Rev Rep, 2011, 7(4): 883-897.
10. Yu HB, Xiong J, Zhang HZ, et al. TGFβ1-transfected tendon stem cells promote tendon fibrosis. J Orthop Surg Res, 2022, 17(1): 358.
11. Cheng B, Ge H, Zhou J, et al. TSG-6 mediates the effect of tendon derived stem cells for rotator cuff healing. Eur Rev Med Pharmacol Sci, 2014, 18(2): 247-251.
12. Shen H, Cheng L, Zheng Q, et al. Scavenging of reactive oxygen species can adjust the differentiation of tendon stem cells and progenitor cells and prevent ectopic calcification in tendinopathy. Acta Biomater, 2022, 152: 440-452.
13. Zhao J, Qiu P, Wang Y, et al. Chitosan-based hydrogel wound dressing: From mechanism to applications, a review. Int J Biol Macromol, 2023, 244: 125250.
14. Guadarrama-Escobar OR, Serrano-Castañeda P, Anguiano-Almazán E, et al. Chitosan nanoparticles as oral drug carriers. Int J Mol Sci, 2023, 24(5): 4289.
15. Jafernik K, Ładniak A, Blicharska E, et al. Chitosan-based nanoparticles as effective drug delivery systems-A review. Molecules, 2023, 28(4): 1963.
16. Li X, Su Z, Shen K, et al. Eugenol-preconditioned mesenchymal stem cell-derived extracellular vesicles promote antioxidant capacity of tendon stem cells in vitro and in vivo. Oxid Med Cell Longev, 2022, 2022: 3945195.
17. Wang Y, Zhao Y, Ma S, et al. Injective programmable proanthocyanidin coordinated zinc-based composite hydrogel for infected bone repair. Adv Healthc Mater, 2023, 27: e202302690.
18. Chen P, Cui L, Fu SC, et al. The 3D-printed PLGA scaffolds loaded with bone marrow-derived mesenchymal stem cells augment the healing of rotator cuff repair in the rabbits. Cell Transplant, 2020, 29: 963689720973647.
19. Rothrauff BB, Smith CA, Ferrer GA, et al. The effect of adipose-derived stem cells on enthesis healing after repair of acute and chronic massive rotator cuff tears in rats. J Shoulder Elbow Surg, 2019, 28(4): 654-664.
20. Cao Y, Yang S, Zhao D, et al. Three-dimensional printed multiphasic scaffolds with stratified cell-laden gelatin methacrylate hydrogels for biomimetic tendon-to-bone interface engineering. J Orthop Translat, 2020, 23: 89-100.
21. Shin MJ, Shim IK, Kim DM, et al. Engineered cell sheets for the effective delivery of adipose-derived stem cells for tendon-to-bone healing. Am J Sports Med, 2020, 48(13): 3347-3358.
22. Liu Q, Hatta T, Qi J, et al. Novel engineered tendon-fibrocartilage-bone composite with cyclic tension for rotator cuff repair. J Tissue Eng Regen Med, 2018, 12(7): 1690-1701.
23. Rui YF, Lui PP, Lee YW, et al. Higher BMP receptor expression and BMP-2-induced osteogenic differentiation in tendon-derived stem cells compared with bone-marrow-derived mesenchymal stem cells. Int Orthop, 2012, 36(5): 1099-1107.
24. Mandalia K, Mousad A, Welborn B, et al. Scaffold- and graft-based biological augmentation of rotator cuff repair: an updated systematic review and meta-analysis of preclinical and clinical studies for 2010-2022. J Shoulder Elbow Surg, 2023, 32(9): 1784-1800.
25. Credille KT, Wang ZRC, Horner NS, et al. Biphasic interpositional allograft for rotator cuff repair augmentation is safe in an ovine model. Arthroscopy, 2023, 39(9): 1983-1997.
26. Abourehab MAS, Pramanik S, Abdelgawad MA, et al. Recent advances of chitosan formulations in biomedical applications. Int J Mol Sci, 2022, 23(18): 10975.
27. Willbold E, Wellmann M, Welke B, et al. Possibilities and limitations of electrospun chitosan-coated polycaprolactone grafts for rotator cuff tear repair. J Tissue Eng Regen Med, 2020, 14(1): 186-197.
28. Niu Y, Wu J, Kang Y, et al. Recent advances of magnetic chitosan hydrogel: Preparation, properties and applications. Int J Biol Macromol, 2023, 247: 125722.

Chinese Journal of Reparative and Reconstructive Surgery

Study on injectable chitosan hydrogel with tendon-derived stem cells for enhancing rotator cuff tendon-to-bone healing

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content