Biopolym. Cell. 2023; 39(2):90-109.
From mesenchymal stem cells to their secretoms as basic components of dermal coverings for the treatment of massive burns
1Papuga O. Ye., 1Macewicz L. L., 1Lukash L. L.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143


In the course of our long term research, we have created new biotechnological products – dermal coverings (dermis equivalents) with the inclusion of human MSCs or their secretomes (cells conditioned mediums which contain a complex of biologically active substances synthesized by them). Preclinical studies on the model animals and clinical trials of new dermal equivalents on the limited contingent of patients with massive burns have been conducted to determine their therapeutic effectiveness as well as safety. We proved at special in vivo experiments that new dermal coverings contribute positively to the healing severe deep burn wounds when applied to the wound surface and don not reveal any toxic property in the studied organisms. The method of obtaining new biotechnological products has been patented.
Keywords: cellular biotechnology, dermal covering or dermis equivalent, mesenchymal stem cell (MSC), conditioned media (CM), secretome, burns


[1] Papuga AYe, Lukash LL. Different types of biotechnological wound coverages created with the application of alive human cells. Biopolym Cell. 2015; 31(2):83-96.
[2] Kosenko OO, Lukash LL, Samchenko YuM, Ruban TA, Ulberg ZR, Lukash SI. Copolymeric hydrogel membranes for immobilization and cultivation of human stem cells. Biopolym Cell. 2006; 22(2):143-8.
[3] Kosenko OO, Lukash LL, Samchenko YuM, Ruban TA, Lukash SI, Ulberg ZR, Galagan NP. Artificial skin equivalent based on copolymeric hydrogel membranes with immobilized human mesenchymal stem cells. Biopolym Cell. 2006; 22(6):446-51.
[4] Ukraine patent for the invention No.82583 "Biocompatible hydrogel of medical prescription and method of its obtaining". Published on April 25, 2008, Bulletin No.1. Samchenko YuM, Lukash LL, Kosenko OO, Ulberg ZR, Ruban TP, Kozynets GP.
[5] Liu P, Deng Z, Han S, Liu T, Wen N, Lu W, Geng X, Huang S, Jin Y. Tissue-engineered skin containing mesenchymal stem cells improves burn wounds. Artif Organs. 2008; 32(12):925-31.
[6] Huang S, Lu G, Wu Y, Jirigala E, Xu Y, Ma K, Fu X. Mesenchymal stem cells delivered in a microsphere-based engineered skin contribute to cutaneous wound healing and sweat gland repair. J Dermatol Sci. 2012; 66(1):29-36.
[7] Chan RK, Zamora DO, Wrice NL, Baer DG, Renz EM, Christy RJ, Natesan S. Development of a vascularized skin construct using adipose-derived stem cells from debrided burned skin. Stem Cells Int. 2012; 2012:841203.
[8] Quan R, Zheng X, Xu S, Zhang L, Yang D. Gelatin-chondroitin-6-sulfate-hyaluronic acid scaffold seeded with vascular endothelial growth factor 165 modified hair follicle stem cells as a three-dimensional skin substitute. Stem Cell Res Ther. 2014; 5(5):118.
[9] Huang S, Wu Y, Gao D, Fu X. Paracrine action of mesenchymal stromal cells delivered by microspheres contributes to cutaneous wound healing and prevents scar formation in mice. Cytotherapy. 2015; 17(7):922-31.
[10] Wu Y, Peng Y, Gao D, Feng C, Yuan X, Li H, Wang Y, Yang L, Huang S, Fu X. Mesenchymal stem cells suppress fibroblast proliferation and reduce skin fibrosis through a TGF-β3-dependent activation. Int J Low Extrem Wounds. 2015; 14(1):50-62.
[11] Zhang B, Wu Y, Mori M, Yoshimura K. Adipose-Derived Stem Cell Conditioned Medium and Wound Healing: A Systematic Review. Tissue Eng Part B Rev. 2022; 28(4):830-47.
[12] Papuga AYe, Samchenko YuM, Ulberg ZR, Ruban TA, Kozinets GP, Lukash LL. The artificial skin equivalent based on acrylic hydrogel with immobilized silver nanoparticles and human cells. Factors in experimental evolution of organisms. 2014; 15:121-4.
[13] Papuga AYe, Samchenko YuM, Suchorada OM, Ruban TA, Kozinets GP, Zenich AV, Uvarova IV, Ulberg ZR, Lukash LL. The influence of nanoparticles and metal iones on the survival and proliferation of human stem cells in vitro. Factors in experimental evolution of organisms. 2015; 17:225-9.
[14] Papuga AYe, Macewicz LL, Ruban TA, Lukash LL. Investigation of dermal equivalents efficacy for the treatment of severe burn wounds. Factors in experimental evolution of organisms. 2016; 19:172-5.
[15] Ukraine patent for the invention No.112584 "A method of obtaining a temporary equivalent of the dermal layer of the skin". Published on September 26, 2016, Bulletin No.18/2016. Papuga AYe, Ruban TA, Macewicz LL, Lukash LL, Lukash SI.
[16] Macewicz LL, Papuga AYe, Ruban TA, Lukash LL. Investigation of cell-derived preparations efficacy for the treatment of severe burn wounds. Factors in experimental evolution of organisms. 2017; 20:232-6.
[17] Lukash LL, Iatsishina AP, Kushniruk VO, Pidpala OV. Reprogramming adult somatic human cells in vitro. Factors in experimental evolution of organisms. 2011; 11:493-8.
[18] Akopyan HR, Huleyuk NL, Kushniruk VO, Mykytenko DO, Iatsyshyna AP, Lukash LL. Comparative analysis of the karyotype of new human cell line 4BL at long-term cultivation: Ploidy of the chromosomal set. Cytol Genet. 2013; 47(5);305-17.
[19] Macewicz LL, Papuga AYe, Ruban TA, Beregova TV, Lukash LL. Optimization of containing dermal coverages production for burn wounds treatment at in vivo model. Factors in experimental evolution of organisms. 2018; 22:287-92.
[20] Ukraine patent for the useful model No.127876 "A method of obtaining a temporary plastic wound coverage "gel-cell" for the treatment of burn wounds". Published on 2018, August 27, Bulletin No.16. Papuga AYe, Lukash LL, Beregova TV, Macewicz LL, Ruban TA, Medvedeva NS.
[21] Lukash LL. Cellular biotechnologies for the development of new dermal equivalents for the treatment of massive burns (Transcript of scientific report at the meeting of the Presidium of NAS of Ukraine October 5, 2022). Visnyk of NAS of Ukraine. 2022; (11):54-9.
[22] Ning CC, Logsetty S, Ghughare S, Liu S. Effect of hydrogel grafting, water and surfactant wetting on the adherence of PET wound dressings. Burns. 2014; 40(6):1164-71.
[23] Asghari S, Logsetty S, Liu S. Imparting commercial antimicrobial dressings with low-adherence to burn wounds. Burns. 2016; 42(4):877-83.
[24] Han L, Lu X, Liu K, Wang K, Fang L, Weng LT, Zhang H, Tang Y, Ren F, Zhao C, Sun G, Liang R, Li Z. Mussel-Inspired Adhesive and Tough Hydrogel Based on Nanoclay Confined Dopamine Polymerization. ACS Nano. 2017; 11(3):2561-74.
[25] Chen T, Chen Y, Rehman HU, Chen Z, Yang Z, Wang M, Li H, Liu H. Ultratough, Self-Healing, and Tissue-Adhesive Hydrogel for Wound Dressing. ACS Appl Mater Interfaces. 2018; 10(39):33523-31.
[26] Wu Z, Hong Y. Combination of the Silver-Ethylene Interaction and 3D Printing To Develop Antibacterial Superporous Hydrogels for Wound Management. ACS Appl Mater Interfaces. 2019; 11(37):33734-47.
[27] Peng Y, Wang Z, Zhou Y, Wang F, Zhang S, He D, Deng L. Ferrocene-functionalized hybrid hydrogel dressing with high-adhesion for combating biofilm. Mater Sci Eng C Mater Biol Appl. 2021; 125:112111.
[28] Yang Z, Huang R, Zheng B, Guo W, Li C, He W, Wei Y, Du Y, Wang H, Wu D, Wang H. Highly Stretchable, Adhesive, Biocompatible, and Antibacterial Hydrogel Dressings for Wound Healing. Adv Sci (Weinh). 2021; 8(8):2003627.
[29] Jiang F, Chi Z, Ding Y, Quan M, Tian Y, Shi J, Song F, Liu C. Wound Dressing Hydrogel of Enteromorpha prolifera Polysaccharide-Polyacrylamide Composite: A Facile Transformation of Marine Blooming into Biomedical Material. ACS Appl Mater Interfaces. 2021; 13(12):14530-42.
[30] Glushchenko NN, Bogoslovskaya OA, Olkhovskaya IP. Physical and chemical laws of biological effect of superfine metal powders. Chemical Physics. 2002; 21(4):79-86.
[31] Tian J, Wong KK, Ho CM, Lok CN, Yu WY, Che CM, Chiu JF, Tam PK. Topical delivery of silver nanoparticles promotes wound healing. ChemMedChem. 2007; 2(1):129-36.
[32] Greulich C, Kittler S, Epple M, Muhr G, Köller M. Studies on the biocompatibility and the interaction of silver nanoparticles with human mesenchymal stem cells (hMSCs). Langenbecks Arch Surg. 2009; 394(3):495-502.
[33] Hackenberg S, Scherzed A, Kessler M, Hummel S, Technau A, Froelich K, Ginzkey C, Koehler C, Hagen R, Kleinsasser N. Silver nanoparticles: evaluation of DNA damage, toxicity and functional impairment in human mesenchymal stem cells. Toxicol Lett. 2011; 201(1):27-33.
[34] Walker M, Will I, Pratt A, Chechik V, Genever P, Ungar D. Magnetically Triggered Release of Entrapped Bioactive Proteins from Thermally Responsive Polymer-Coated Iron Oxide Nanoparticles for Stem-Cell Proliferation. ACS Appl Nano Mater. 2020; 3(6):5008-13.
[35] Tran N, Mir A, Mallik D, Sinha A, Nayar S, Webster TJ. Bactericidal effect of iron oxide nanoparticles on Staphylococcus aureus. Int J Nanomedicine. 2010; 5:277-83.
[36] Cheng FY, Su CH, Yang YS, Yeh CS, Tsai CY, Wu CL, Wu MT, Shieh DB. Characterization of aqueous dispersions of Fe(3)O(4) nanoparticles and their biomedical applications. Biomaterials. 2005; 26(7):729-38.
[37] Brunner TJ, Wick P, Manser P, Spohn P, Grass RN, Limbach LK, Bruinink A, Stark WJ. In vitro cytotoxicity of oxide nanoparticles: comparison to asbestos, silica, and the effect of particle solubility. Environ Sci Technol. 2006; 40(14):4374-81.
[38] Kukharenko O, Bardeau J-F, Zaets I, Ovcharenko L, Tarasyuk O, Porhyn S, Mischenko I, Vovk A, Rogalsky S, Kozyrovska N. Promising low cost antimicrobial composite material based on bacterial cellulose and polyhexamethylene guanidine hydrochloride. European Polymer Journal. 2014; 60:247-54.
[39] Pianigiani E, Andreassi A, Taddeucci P, Alessandrini C, Fimiani M, Andreassi L. A new model for studying differentiation and growth of epidermal cultures on hyaluronan-based carrier. Biomaterials. 1999; 20(18):1689-94.
[40] Sharif S, Ai J, Azami M, Verdi J, Atlasi MA, Shirian S, Samadikuchaksaraei A. Collagen-coated nano-electrospun PCL seeded with human endometrial stem cells for skin tissue engineering applications. J Biomed Mater Res B Appl Biomater. 2018; 106(4):1578-86.
[41] Ababzadeh S, Farzin A, Goodarzi A, Karimi R, Sagharjoghi Farahani M, Eslami Farsani M, Gharibzad K, Zahiri M, Ai J. High porous electrospun poly(ε-caprolactone)/gelatin/MgO scaffolds preseeded with endometrial stem cells promote tissue regeneration in full-thickness skin wounds: An in vivo study. J Biomed Mater Res B Appl Biomater. 2020; 108(7):2961-70.
[42] Roseeuw DI, de Coninck AL, Vanpée ES, Delaey BM, Verbeken GL, Draye JP. Healing of full-thickness wounds treated with lyophilized cultured keratinocyte cell lysate in genetically diabetic mice. Wound Repair Regen. 1995; 3(4):500-5.
[43] Akbari A, Li Y, Kilani RT, Ghahary A. Red blood cell lysate modulates the expression of extracellular matrix proteins in dermal fibroblasts. Mol Cell Biochem. 2012; 370(1-2):79-88.
[44] Hur W, Lee HY, Min HS, Wufuer M, Lee CW, Hur JA, Kim SH, Kim BK, Choi TH. Regeneration of full-thickness skin defects by differentiated adipose-derived stem cells into fibroblast-like cells by fibroblast-conditioned medium. Stem Cell Res Ther. 2017;8(1):92.
[45] Maarof M, Lokanathan Y, Ruszymah HI, Saim A, Chowdhury SR. Proteomic Analysis of Human Dermal Fibroblast Conditioned Medium (DFCM). Protein J. 2018; 37(6):589-607.
[46] Maarof M, Mh Busra MF, Lokanathan Y, Bt Hj Idrus R, Rajab NF, Chowdhury SR. Safety and efficacy of dermal fibroblast conditioned medium (DFCM) fortified collagen hydrogel as acellular 3D skin patch. Drug Deliv Transl Res. 2019; 9(1):144-61.
[47] Maarof M, Mohd Nadzir M, Sin Mun L, Fauzi MB, Chowdhury SR, Idrus RBH, Lokanathan Y. Hybrid Collagen Hydrogel/Chondroitin-4-Sulphate Fortified with Dermal Fibroblast Conditioned Medium for Skin Therapeutic Application. Polymers (Basel). 2021; 13(4):508.
[48] Shohara R, Yamamoto A, Takikawa S, Iwase A, Hibi H, Kikkawa F, Ueda M. Mesenchymal stromal cells of human umbilical cord Wharton's jelly accelerate wound healing by paracrine mechanisms. Cytotherapy. 2012; 14(10):1171-81.
[49] Peng Y, Xuan M, Zou J, Liu H, Zhuo Z, Wan Y, Cheng B. Freeze-dried rat bone marrow mesenchymal stem cell paracrine factors: a simplified novel material for skin wound therapy. Tissue Eng Part A. 2015; 21(5-6):1036-46.
[50] Tutuianu R, Rosca AM, Albu Kaya MG, Pruna V, Neagu TP, Lascar I, Simionescu M, Titorencu I. Mesenchymal stromal cell-derived factors promote the colonization of collagen 3D scaffolds with human skin cells. J Cell Mol Med. 2020; 24(17):9692-704.
[51] Krasnodembskaya A, Song Y, Fang X, Gupta N, Serikov V, Lee JW, Matthay MA. Antibacterial effect of human mesenchymal stem cells is mediated in part from secretion of the antimicrobial peptide LL-37. Stem Cells. 2010; 28(12):2229-38.
[52] Walter MN, Wright KT, Fuller HR, MacNeil S, Johnson WE. Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays. Exp Cell Res. 2010; 316(7):1271-81.
[53] Kaita Y, Tarui T, Yoshino H, Matsuda T, Yamaguchi Y, Nakagawa T, Asahi M, Ii M. Sufficient therapeutic effect of cryopreserved frozen adipose-derived regenerative cells on burn wounds. Regen Ther. 2019; 10:92-103.
[54] Yang J, Chen Z, Pan D, Li H, Shen J. Umbilical Cord-Derived Mesenchymal Stem Cell-Derived Exosomes Combined Pluronic F127 Hydrogel Promote Chronic Diabetic Wound Healing and Complete Skin Regeneration. Int J Nanomedicine. 2020; 15:5911-26.
[55] Kim MH, Wu WH, Choi JH, Kim JH, Hong SH, Jun JH, Ko Y, Lee JH. Conditioned medium from the three-dimensional culture of human umbilical cord perivascular cells accelerate the migration and proliferation of human keratinocyte and fibroblast. J Biomater Sci Polym Ed. 2018; 29(7-9):1066-80.
[56] Li M, Luan F, Zhao Y, Hao H, Liu J, Dong L, Fu X, Han W. Mesenchymal stem cell-conditioned medium accelerates wound healing with fewer scars. Int Wound J. 2017; 14(1):64-73.
[57] Zhang B, Wu X, Zhang X, Sun Y, Yan Y, Shi H, Zhu Y, Wu L, Pan Z, Zhu W, Qian H, Xu W. Human umbilical cord mesenchymal stem cell exosomes enhance angiogenesis through the Wnt4/β-catenin pathway. Stem Cells Transl Med. 2015; 4(5):513-22.
[58] Komaki M, Numata Y, Morioka C, Honda I, Tooi M, Yokoyama N, Ayame H, Iwasaki K, Taki A, Oshima N, Morita I. Exosomes of human placenta-derived mesenchymal stem cells stimulate angiogenesis. Stem Cell Res Ther. 2017; 8(1):219.
[59] Vizoso FJ, Eiro N, Cid S, Schneider J, Perez-Fernandez R. Mesenchymal Stem Cell Secretome: Toward Cell-Free Therapeutic Strategies in Regenerative Medicine. Int J Mol Sci. 2017; 18(9):1852.
[60] Ferreira JR, Teixeira GQ, Santos SG, Barbosa MA, Almeida-Porada G, Gonçalves RM. Mesenchymal Stromal Cell Secretome: Influencing Therapeutic Potential by Cellular Pre-conditioning. Front Immunol. 2018; 9:2837.
[61] Shablii VA, Kuchma MD, Kyryk VM, Onishchenko AN, Lukash LL, Lobitseva GS. Cryopreservation human placental tissue as source of hematopoietic and mesenchymal stem cells. Cell Transplantation and Tissue Engineering. 2012; 7(1):54-62.
[62] Shablii VA, Kuchma MD, Kyryk VM, Onishchenko AN, Areshkov PO, Skrypnyk N, Lukash LL, Lobyntseva GS. Characteristics of placental multipotent mesenchymal stromal stem cells. Cell Transplantation and Tissue Engineering. 2012; 7(4):55-61.
[63] Shablii VA, Kuchma MD, Kyryk VM, Svitina HM, Shablii YM, Lukash LL, Lobintseva GS. Mesenchymal and trophoblast immunophenotype of multipotent stromal cells from human placenta. Biopolym Cell. 2014; 30(2):118-21.
[64] Muhammad SA, Nordin N, Fakurazi S. Regenerative potential of secretome from dental stem cells: a systematic review of preclinical studies. Rev Neurosci. 2018; 29(3):321-32.
[65] Fui LW, Lok MPW, Govindasamy V, Yong TK, Lek TK, Das AK. Understanding the multifaceted mechanisms of diabetic wound healing and therapeutic application of stem cells conditioned medium in the healing process. J Tissue Eng Regen Med. 2019; 13(12):2218-33.
[66] Md Fadilah NI, Mohd Abdul Kader Jailani MS, Badrul Hisham MAI, Sunthar Raj N, Shamsuddin SA, Ng MH, Fauzi MB, Maarof M. Cell secretomes for wound healing and tissue regeneration: Next generation acellular based tissue engineered products. J Tissue Eng. 2022; 13:20417314221114273.
[67] Abdul Kareem N, Aijaz A, Jeschke MG. Stem Cell Therapy for Burns: Story so Far. Biologics. 2021; 15:379-97.
[68] Damayanti RH, Rusdiana T, Wathoni N. Mesenchymal Stem Cell Secretome for Dermatology Application: A Review. Clin Cosmet Investig Dermatol. 2021; 14:1401-12.