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The Biocompatibility of Multi-Source Stem Cells and Gelatin-Carboxymethyl Chitosan-Sodium Alginate Hybrid Biomaterials



Nowadays, biological tissue engineering is a growing field of research. Biocompatibility is a key indicator for measuring tissue engineering biomaterials, which is of great significance for the replacement and repair of damaged tissues.


In this study, using gelatin, carboxymethyl chitosan, and sodium alginate, a tissue engineering material scaffold that can carry cells was successfully prepared. The material was characterized by Fourier transforms infrared spectroscopy. In addition, the prepared scaffolds have physicochemical properties, such as swelling ratio, biodegradability. we observed the biocompatibility of the hydrogel to different adult stem cells (BMSCs and ADSCs) in vivo and in vitro. Adult stem cells were planted on gelatin-carboxymethyl chitosan-sodium alginate (Gel/SA/CMCS) hydrogels for 7 days in vitro, and the survival of stem cells in vitro was observed by live/died staining. Gel/SA/CMCS hydrogels loaded with stem cells were subcutaneously transplanted into nude mice for 14 days of in vivo culture observation. The survival of adult stem cells was observed by staining for stem cell surface markers (CD29, CD90) and Ki67.


The scaffolds had a microporous structure with an appropriate pore size (about 80 μm). Live/died staining showed that adult stem cells could stably survive in Gel/SA/CMCS hydrogels for at least 7 days. After 14 days of culture in nude mice, Ki67 staining showed that the stem cells supported by Gel/SA/CMCS hydrogel still had high proliferation activity.


Gel/SA/CMCSs hydrogel has a stable interpenetrating porous structure, suitable swelling performance and degradation rate, can promote and support the survival of adult stem cells in vivo and in vitro, and has good biocompatibility. Therefore, Gel/SA/CMCS hydrogel is a strong candidate for biological tissue engineering materials.

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  1. Zurina IM, Presniakova VS, Butnaru DV, Svistunov AA, Timashev PS, Rochev YA. Tissue engineering using a combined cell sheet technology and scaffolding approach. Acta Biomater. 2020;113:63–83.

    CAS  Article  Google Scholar 

  2. Berthiaume F, Maguire TJ, Yarmush ML. Tissue engineering and regenerative medicine: history, progress, and challenges. Annu Rev Chem Biomol Eng. 2011;2:403–30.

    Article  Google Scholar 

  3. Wang Y, Kim HJ, Vunjak-Novakovic G, Kaplan DL. Stem cell-based tissue engineering with silk biomaterials. Biomaterials. 2006;27:6064–82.

    CAS  Article  Google Scholar 

  4. He Y, Zhao W, Dong Z, Ji Y, Li M, Hao Y, et al. A biodegradable antibacterial alginate/carboxymethyl chitosan/Kangfuxin sponges for promoting blood coagulation and full-thickness wound healing. Int J Biol Macromol. 2021;167:182–92.

    CAS  Article  Google Scholar 

  5. Huang J, Fu H, Wang Z, Meng Q, Liu S, Wang H, et al. BMSCs-laden gelatin/sodium alginate/carboxymethyl chitosan hydrogel for 3D bioprinting. RSC Adv. 2016;6:108423–30.

    CAS  Article  Google Scholar 

  6. Ansari S, Diniz IM, Chen C, Sarrion P, Tamayol A, Wu BM, et al. Human periodontal ligament- and gingiva-derived mesenchymal stem cells promote nerve regeneration when encapsulated in alginate/hyaluronic acid 3D scaffold. Adv Healthc Mater. 2017;6:1700670.

  7. He Y, Li Y, Sun Y, Zhao S, Feng M, Xu G, et al. A double-network polysaccharide-based composite hydrogel for skin wound healing. Carbohydr Polym. 2021;261:117870.

    CAS  Article  Google Scholar 

  8. Yang X, Yang H, Jiang X, Yang B, Zhu K, Lai NC, et al. Injectable chitin hydrogels with self-healing property and biodegradability as stem cell carriers. Carbohydr Polym. 2021;256:117574.

    CAS  Article  Google Scholar 

  9. Liu X, Hao M, Chen Z, Zhang T, Huang J, Dai J, et al. 3D bioprinted neural tissue constructs for spinal cord injury repair. Biomaterials. 2021;272:120771.

    CAS  Article  Google Scholar 

  10. Hassanzadeh P, Atyabi F, Dinarvand R. Tissue engineering: Still facing a long way ahead. J Control Release. 2018;279:181–97.

    CAS  Article  Google Scholar 

  11. Shafiee A, Atala A. Tissue engineering: Toward a new era of medicine. Annu Rev Med. 2017;68:29–40.

    CAS  Article  Google Scholar 

  12. Gu Z, Huang K, Luo Y, Zhang L, Kuang T, Chen Z, et al. Double network hydrogel for tissue engineering. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2018;10:e1520.

    Article  Google Scholar 

  13. Long Y, Yan L, Dai H, Yang D, Wu X, Dong X, et al. Enhanced proliferation and differentiation of neural stem cells by peptide-containing temperature-sensitive hydrogel scaffold. Mater Sci Eng C Mater Biol Appl. 2020;116:111258.

    CAS  Article  Google Scholar 

  14. Bakhshandeh B, Zarrintaj P, Oftadeh MO, Keramati F, Fouladiha H, Sohrabi-Jahromi S, et al. Tissue engineering; strategies, tissues, and biomaterials. Biotechnol Genet Eng Rev. 2017;33:144–72.

    CAS  Article  Google Scholar 

  15. Huang GY, Zhou LH, Zhang QC, Chen YM, Sun W, Xu F, et al. Microfluidic hydrogels for tissue engineering. Biofabrication. 2011;3:012001.

    Article  Google Scholar 

  16. Hu Y, Dan W, Xiong S, Kang Y, Dhinakar A, Wu J, et al. Development of collagen/polydopamine complexed matrix as mechanically enhanced and highly biocompatible semi-natural tissue engineering scaffold. Acta Biomater. 2017;47:135–48.

    CAS  Article  Google Scholar 

  17. Basu A, Kunduru KR, Abtew E, Domb AJ. Polysaccharide-Based Conjugates for Biomedical Applications. Bioconjug Chem. 2015;26:1396–412.

    CAS  Article  Google Scholar 

  18. Varaprasad K, Jayaramudu T, Kanikireddy V, Toro C, Sadiku ER. Alginate-based composite materials for wound dressing application: A mini review. Carbohydr Polym. 2020;236:116025.

    CAS  Article  Google Scholar 

  19. Wang YL, Zhou YN, Li XY, Huang J, Wahid F, Zhong C. Continuous production of antibacterial carboxymethyl chitosan-zinc supramolecular hydrogel fiber using a double-syringe injection device. Int J Biol Macromol. 2020;156:252–61.

    CAS  Article  Google Scholar 

  20. Wang D, Zhang N, Meng G, He J, Wu F. The effect of form of carboxymethyl-chitosan dressings on biological properties in wound healing. Colloids Surf B Biointerfaces. 2020;194:111191.

    CAS  Article  Google Scholar 

  21. Yan T, Hui W, Zhu S, He J, Liu Z, Cheng J. Carboxymethyl chitosan based redox-responsive micelle for near-infrared fluorescence image-guided photo-chemotherapy of liver cancer. Carbohydr Polym. 2021;253:117284.

    CAS  Article  Google Scholar 

  22. Jiang Z, Wang S, Hou J, Chi J, Wang S, Shao K, et al. Effects of carboxymethyl chitosan oligosaccharide on regulating immunologic function and inhibiting tumor growth. Carbohydr Polym. 2020;250:116994.

    CAS  Article  Google Scholar 

  23. Aljohani W, Ullah MW, Zhang X, Yang G. Bioprinting and its applications in tissue engineering and regenerative medicine. Int J Biol Macromol. 2018;107:261–75.

    CAS  Article  Google Scholar 

  24. Vorwald CE, Gonzalez-Fernandez T, Joshee S, Sikorski P, Leach JK. Tunable fibrin-alginate interpenetrating network hydrogels to support cell spreading and network formation. Acta Biomater. 2020;108:142–52.

    CAS  Article  Google Scholar 

  25. Liu Q, Wang J, Chen Y, Zhang Z, Saunders L, Schipani E, et al. Suppressing mesenchymal stem cell hypertrophy and endochondral ossification in 3D cartilage regeneration with nanofibrous poly(l-lactic acid) scaffold and matrilin-3. Acta Biomater. 2018;76:29–38.

    CAS  Article  Google Scholar 

  26. Teekakirikul P, Zhu W, Huang HC, Fung E. Hypertrophic cardiomyopathy: An overview of genetics and management. Biomolecules. 2019;9:878.

  27. Craciun AM, Mititelu Tartau L, Pinteala M, Marin L. Nitrosalicyl-imine-chitosan hydrogels based drug delivery systems for long term sustained release in local therapy. J Colloid Interface Sci. 2019;536:196–207.

  28. Echave MC, Saenz delBurgo L, Pedraz JL, Orive G. Gelatin as biomaterial for tissue engineering. Curr Pharm Des. 2017;23:3567–84.

    CAS  Article  Google Scholar 

  29. Wang S, Guan S, Li W, Ge D, Xu J, Sun C, et al. 3D culture of neural stem cells within conductive PEDOT layer-assembled chitosan/gelatin scaffolds for neural tissue engineering. Mater Sci Eng C Mater Biol Appl. 2018;93:890–901.

    CAS  Article  Google Scholar 

  30. Cheng Y, Hu Z, Zhao Y, Zou Z, Lu S, Zhang B, et al. Sponges of carboxymethyl chitosan grafted with collagen peptides for wound healing. Int J Mol Sci. 2019;20:3890.

  31. Yu H, Zhang X, Song W, Pan T, Wang H, Ning T, et al. Effects of 3-dimensional bioprinting alginate/gelatin hydrogel scaffold extract on proliferation and differentiation of human dental pulp stem cells. J Endod. 2019;45:706–15.

    Article  Google Scholar 

  32. Wang J, Zhou L, Sun Q, Cai H, Tan WS. Porous chitosan derivative scaffolds affect proliferation and osteogenesis of mesenchymal stem cell via reducing intracellular ROS. Carbohydr Polym. 2020;237:116108.

    CAS  Article  Google Scholar 

  33. Wang K, Nune KC, Misra RD. The functional response of alginate-gelatin-nanocrystalline cellulose injectable hydrogels toward delivery of cells and bioactive molecules. Acta Biomater. 2016;36:143–51.

    CAS  Article  Google Scholar 

  34. An S, Ling J, Gao Y, Xiao Y. Effects of varied ionic calcium and phosphate on the proliferation, osteogenic differentiation and mineralization of human periodontal ligament cells in vitro. J Periodontal Res. 2012;47:374–82.

    CAS  Article  Google Scholar 

  35. Zimet P, Mombru AW, Mombru D, Castro A, Villanueva JP, Pardo H, Rufo C. Physico-chemical and antilisterial properties of nisin-incorporated chitosan/carboxymethyl chitosan films. Carbohydr Polym. 2019;219:334–43.

    CAS  Article  Google Scholar 

  36. Shariatinia Z. Carboxymethyl chitosan: Properties and biomedical applications. Int J Biol Macromol. 2018;120:1406–19.

    CAS  Article  Google Scholar 

  37. Wahid F, Yin JJ, Xue DD, Xue H, Lu YS, Zhong C, et al. Synthesis and characterization of antibacterial carboxymethyl Chitosan/ZnO nanocomposite hydrogels. Int J Biol Macromol. 2016;88:273–9.

    CAS  Article  Google Scholar 

  38. Zhao X, Li P, Guo B, Ma PX. Antibacterial and conductive injectable hydrogels based on quaternized chitosan-graft-polyaniline/oxidized dextran for tissue engineering. Acta Biomater. 2015;26:236–48.

    CAS  Article  Google Scholar 

  39. Periayah MH, Halim AS, Mat Saad AZ. Mechanism action of platelets and crucial blood coagulation pathways in hemostasis. Int J Hematol Oncol Stem Cell Res. 2017;11:319–27.

    PubMed  PubMed Central  Google Scholar 

  40. Bakshi PS, Selvakumar D, Kadirvelu K, Kumar NS. Chitosan as an environment friendly biomaterial - a review on recent modifications and applications. Int J Biol Macromol. 2020;150:1072–83.

    CAS  Article  Google Scholar 

  41. Chen Y, Wu L, Li P, Hao X, Yang X, Xi G, et al. Polysaccharide based hemostatic strategy for ultrarapid hemostasis. Macromol Biosci. 2020;20:e1900370.

    Article  Google Scholar 

  42. Liu Y, Sui Y, Liu C, Liu C, Wu M, Li B, et al. A physically crosslinked polydopamine/nanocellulose hydrogel as potential versatile vehicles for drug delivery and wound healing. Carbohydr Polym. 2018;188:27–36.

    CAS  Article  Google Scholar 

  43. Li S, Dong S, Xu W, Tu S, Yan L, Zhao C, et al. Antibacterial hydrogels. Adv Sci (Weinh). 2018;5:1700527.

  44. Mao C, Xiang Y, Liu X, Cui Z, Yang X, Li Z, et al. Repeatable photodynamic therapy with triggered signaling pathways of fibroblast cell proliferation and differentiation to promote bacteria-accompanied wound healing. ACS Nano. 2018;12:1747–59.

    CAS  Article  Google Scholar 

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This work was supported by Shandong Provincial Natural Science Foundation of China (No. ZR2020MH070 and No. ZR2020MH078), Shandong Province Medicine and Health Science and Technology Development Plan Project (No. 2019WS368), and Research Support Foundation of Jining Medical University (No. JYFC2018FKJ009 and No. JYFC2018KJ004). Xinzhe Wang: Methodology, Investigation, Formal analysis, Writing-original draft. Siqi Li: Investigation, Data curation. Honglian Yu: Conceptualization, Formal analysis, Writing-review & editing, Supervision, Project administration. Jianzhi Lv: Validation, Data curation. Minglun Fan: Investigation. Ximing Wang: Validation. Xin Wang: Validation. Yanting Liang: Validation. Lingna Mao: Validation. Zhankui Zhao: Supervision, Writing—review & editing.

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Correspondence to Zhankui Zhao.

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All animal experiments were conducted in the Central Laboratory of Affiliated Hospital of Jining Medical University, according to the guidelines of the Institutional Animal Care and Use Committee and the Declaration of Helsinki.

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Wang, X., Li, S., Yu, H. et al. The Biocompatibility of Multi-Source Stem Cells and Gelatin-Carboxymethyl Chitosan-Sodium Alginate Hybrid Biomaterials. Tissue Eng Regen Med 19, 491–503 (2022).

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  • Adult stem cells
  • Sodium alginate
  • Carboxymethyl chitosan
  • Histocompatibility