Skip to main content Skip to main navigation menu Skip to site footer

Composite characterization of freeze-dried human amnion membrane and human adipose tissue derived stromal cells for soft tissue engineering

  • Yusuf Rizal ,
  • Mouli Edward ,
  • Tri Wahyu Martanto ,
  • Heri Suroto ,


Background: Soft tissue engineering has widely explored freeze-dried human amniotic membrane (FdHAM) and human adipose-derived stromal cells (hADSC) as separate components. In this study, the composite of FdHAM and hADSC combination was investigated regarding cell morphology and density, constituent components, and cytotoxicity evaluation.

Methods: In vitro experimental laboratory research was conducted to analysis of the FdHAM and hADSC composites. Evaluation of cell viability and CD105 for hADSC, evaluation of pore diameter for amniotic membrane and assessment of composite cell morphology, constituent components, evaluation of cytotoxicity, and cell density was conducted.

Results: Our morphology data indicated that clusters of hADSC with homogenous distribution on FdHAM. The viability test showed that the hADSCs were still dominantly alive with an average percentage of 69.3% with the mean cell concentration being 1.53 x 106/mL. The average FdHAM pore diameter was 1.90±0.30 µM. Our data suggested that FdHAM was nontoxic with good cell survivability after exposure to FdHAM.

Conclusions: The structure of FdHAM and hADSC has excellent basic properties and therefore suitable for use as carriers of hADSC or in soft tissue engineering. Compared to the available literature, the characteristic of the FdHAM and hADSC composite is ideal and may be used as an end-product for use in humans.


  1. Dadkhah Tehrani F, Firouzeh A, Shabani I, Shabani A. A Review on Modifications of Amniotic Membrane for Biomedical Applications. Front Bioeng Biotechnol. 2021;8. Available from:
  2. Suroto H, Aryawan DM, Prakoeswa CA. The Influence of the Preservation Method and Gamma Irradiation Sterilization on TGF-β and bFGF Levels in Freeze-Dried Amnion Membrane (FD-AM) and Amnion Sponge. Int J Biomater. 2021;2021:6685225. Available from:
  3. Laloze J, Fiévet L, Desmoulière A. Adipose-Derived Mesenchymal Stromal Cells in Regenerative Medicine: State of Play, Current Clinical Trials, and Future Prospects. Adv wound care. 2020/06/02. 2021;10(1):24–48. Available from:
  4. Jancuska J, Matthews J, Miller T, Kluczynski MA, Bisson LJ. A Systematic Summary of Systematic Reviews on the Topic of the Rotator Cuff. Orthop J Sport Med. 2018;6(9):2325967118797891–2325967118797891. Available from:
  5. Hilmy N, Yusof N, Nather A. Human Amniotic Membrane [Internet]. WORLD SCIENTIFIC; 2017. Available from:
  6. Kikuchi-Taura A, Taguchi A, Kanda T, Inoue T, Kasahara Y, Hirose H, et al. Human umbilical cord provides a significant source of unexpanded mesenchymal stromal cells. Cytotherapy. 2012;14(4):441–50. Available from:
  7. Pan H-C, Yang D-Y, Chiu Y-T, Lai S-Z, Wang Y-C, Chang M-H, et al. Enhanced regeneration in injured sciatic nerve by human amniotic mesenchymal stem cell. J Clin Neurosci. 2006;13(5):570–5. Available from:
  8. Wei JP, Zhang TS, Kawa S, Aizawa T, Ota M, Akaike T, et al. Human Amnion-Isolated Cells Normalize Blood Glucose in Streptozotocin-Induced Diabetic Mice. Cell Transplant. 2003;12(5):545–52. Available from:
  9. Kim J, Jeong SY, Ju YM, Yoo JJ, Smith TL, Khang G, et al. In vitro osteogenic differentiation of human amniotic fluid-derived stem cells on a poly(lactide- co -glycolide) (PLGA)–bladder submucosa matrix (BSM) composite scaffold for bone tissue engineering. Biomed Mater. 2013;8(1):14107. Available from:
  10. Lee SJ, Lim GJ, Lee J-W, Atala A, Yoo JJ. In vitro evaluation of a poly(lactide-co-glycolide)–collagen composite scaffold for bone regeneration. Biomaterials. 2006;27(18):3466–72. Available from:
  11. Leong WS, Tay CY, Yu H, Li A, Wu SC, Duc D-H, et al. Thickness sensing of hMSCs on collagen gel directs stem cell fate. Biochem Biophys Res Commun. 2010;401(2):287–92. Available from:
  12. Vissers CAB, Harvestine JN, Leach JK. Pore size regulates mesenchymal stem cell response to Bioglass-loaded composite scaffolds. J Mater Chem B. 2015;3(44):8650–8. Available from:
  13. Eivazzadeh-Keihan R, Maleki A, de la Guardia M, Bani MS, Chenab KK, Pashazadeh-Panahi P, et al. Carbon based nanomaterials for tissue engineering of bone: Building new bone on small black scaffolds: A review. J Adv Res. 2019;18:185–201. Available from:
  14. Jiwanti PK, Wardhana BY, Sutanto LG, Dewi DMM, Putri IZD, Savitri INI. Recent Development of Nano-Carbon Material in Pharmaceutical Application: A Review. Molecules. 2022;27(21):7578. Available from:
  15. Zubaidah N. The citotoxicity of calcium hydroxide intracanal dressing by MTT assay. Dent J (Majalah Kedokt Gigi). 2007;40(4):157. Available from:
  16. Kusumaningsih T, Pratiwi FD, Sunariani J. Cytotoxicity Test of Amethyst Root Extract (Datura metel L.) on Fibroblast Cell Using MTT Assay. Oral Biology Dental Journal. 2015;7(1):38-43.
  17. Ghasemi M, Turnbull T, Sebastian S, Kempson I. The MTT Assay: Utility, Limitations, Pitfalls, and Interpretation in Bulk and Single-Cell Analysis. Int J Mol Sci. 2021;22(23):12827. Available from:

How to Cite

Rizal, Y., Edward, M. ., Martanto, T. W. ., & Suroto, H. . (2023). Composite characterization of freeze-dried human amnion membrane and human adipose tissue derived stromal cells for soft tissue engineering. Bali Medical Journal, 12(2), 1543–1548.