A pilot study to demonstrate the paracrine effect of equine, adult allogenic mesenchymal stem cells in vitro, with a potential for healing of experimentally-created, equine thoracic wounds in vivo.
Abstract: Regenerative biological therapies using mesenchymal stem cells (MSCs) are being studied and used extensively in equine veterinary medicine. One of the important properties of MSCs is the cells' reparative effect, which is brought about by paracrine signaling, which results in the release of biologically active molecules, which in turn, can affect cellular migration and proliferation, thus a huge potential in wound healing. The objective of the current study was to demonstrate the in vitro and in vivo potentials of equine allogenic bone marrow-derived MSCs for wound healing. Equine bone marrow-derived MSCs from one allogenic donor horse were used. Equine MSCs were previously characterized for their in vitro proliferation, expression of cluster-of-differentiation markers, and trilineage differentiation. MSCs were first evaluated for their migration using an in vitro wound healing scratch assay, and subsequently, the conditioned medium was evaluated for their effect on human fibroblast proliferation. Subsequently, allogenic cells were intradermally injected into full-thickness, cutaneous thoracic wounds of 4 horses. Wound healing was assessed by using 3-D digital imaging and by measuring mRNA expression of pro-and anti-inflammatory markers for 30 days. Using human fibroblasts in an in vitro wound healing assay, we demonstrate a significantly higher healing in the presence of conditioned medium collected from proliferating MSCs than in the presence of medium containing fetal bovine serum. The in vitro effect of MSCs did not translate into a detectable effect in vivo. Nonetheless, we proved that molecularly characterized equine allogenic MSCs do not illicit an immunologic response. Investigations using MSCs derived from other sources (adipose tissue, umbilical cord), or a higher number of MSCs or a compromised animal model may be required to prove the efficacy of equine MSCs in wound healing in vivo.
Copyright © 2022 Caruso, Shuttle, Amelse, Elkhenany, Schumacher and Dhar.
Publication Date: 2022-11-14 PubMed ID: 36452146PubMed Central: PMC9702339DOI: 10.3389/fvets.2022.1011905Google Scholar: Lookup
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Summary
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The research paper pertains to a study exploring the use of adult allogenic mesenchymal stem cells from horses to repair thoracic wounds via paracrine signaling. Researchers sought to demonstrate the effectiveness of this type of regenerative therapy both in vitro (observed under controlled lab conditions) and in vivo (observing effects directly on the living organism).
Research Methodology and Objectives
- The primary goal was to demonstrate the in vitro and in vivo potentials of equine allogenic bone marrow-derived MSCs for wound healing.
- The scientists used MSCs from the bone marrow of one allogenic donor horse for this pilot study.
- The chosen equine MSCs had been previously characterized for their proliferation, expression of cluster-of-differentiation markers, and trilineage differentiation.
- They first conducted an in vitro wound healing scratch assay to evaluate the migration of the MSCs.
- The researchers then examined the effect of the MSCs’ conditioned medium on human fibroblast proliferation.
- Afterwards, they injected allogenic cells intradermally into full-thickness, cutaneous thoracic wounds of 4 horses and assessed the wound healing over a period of 30 days by using 3-D digital imaging and measuring mRNA expression of pro-and anti-inflammatory markers.
Research Findings
- Findings from the in vitro wound healing assay using human fibroblasts signified a significantly higher rate of healing when using the conditioned medium collected from proliferating MSCs compared to using medium containing fetal bovine serum.
- However, this in vitro healing effect of MSCs did not translate into a clear and detectable effect in vivo.
- Despite this, they achieved an important scientific understanding that molecularly characterized, equine allogenic MSCs do not trigger an immunologic response.
- This research laid groundwork for further exploration using MSCs derived from alternative sources like adipose tissue, umbilical cord or increasing the number of MSCs for possible improved efficacy in wound healing.
- Consideration for a compromised animal model was suggested as a next step to test and verify the efficacy of equine MSCs in wound healing.
Cite This Article
APA
Caruso M, Shuttle S, Amelse L, Elkhenany H, Schumacher J, Dhar MS.
(2022).
A pilot study to demonstrate the paracrine effect of equine, adult allogenic mesenchymal stem cells in vitro, with a potential for healing of experimentally-created, equine thoracic wounds in vivo.
Front Vet Sci, 9, 1011905.
https://doi.org/10.3389/fvets.2022.1011905 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Knoxville, TN, United States.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Knoxville, TN, United States.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Knoxville, TN, United States.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Knoxville, TN, United States.
- Department of Surgery, Faculty of Veterinary Medicine, Alexandria University, Alexandria, Egypt.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Knoxville, TN, United States.
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Knoxville, TN, United States.
- Department of Surgery, Faculty of Veterinary Medicine, Alexandria University, Alexandria, Egypt.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
References
This article includes 40 references
- Fu X, Liu G, Halim A, Ju Y, Luo Q, Song G. Mesenchymal stem cell migration and tissue repair. Cells (2019) 8:784.
- Caplan AI. Mesenchymal stem cells: Time to change the name. Stem Cells Transl Med (2017) 6:1445–51.
- Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science (1997) 276:71–4.
- Fathke C, Wilson L, Hutter J, Kapoor V, Smith A, Hocking A. Contribution of bone marrow-derived cells to skin: collagen deposition and wound repair. Stem Cells (2004) 22:812–22.
- Sorrell JM, Caplan AI. Topical delivery of mesenchymal stem cells and their function in wounds. Stem Cell Res Ther (2010) 1:6.
- Singer AJ, Clark RA. Cutaneous wound healing. N Engl J Med (1999) 341:738–46.
- Wu Y, Chen L, Scott PG, Tredget EE. Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells (2007) 25:2648–59.
- Arno AI, Amini-Nik S, Blit PH, Al-Shehab M, Belo C, Herer E. Human Wharton's jelly-mesenchymal stem cells promote skin wound healing through paracrine signaling. Stem Cell Res Therapy (2014) 5:13.
- Nakagawa H, Akita S, Fukui M, Akino K. Human mesenchymal stem cells successfully improve skin-substitute wound healing. Br J Dermatol (2005) 153:29–36.
- Javazon EH, Keswani SG, Badillo AT, Crombleholme TM, Zoltick PW, Radu AP. Enhanced epithelial gap closure and increased angiogenesis in wounds of diabetic mice treated with adult murine bone marrow stromal progenitor cells. Wound Rep Reg (2007) 15:350–9.
- Hocking AM, Gibran NS. Mesenchymal stem cells: paracrine signaling and differentiation during cutaneous wound repair. Exp Cell Res (2010) 316:2213–9.
- Motegi SI, Ishikawa O. Mesenchymal stem cells: the roles and functions in cutaneous wound healing and tumor growth. J Dermatol Sci (2017) 86:83–9.
- Herdrich BJ, Lind RC, Liechty KW. Multipotent adult progenitor cells: their role in wound healing and the treatment of dermal wounds. Cytotherapy (2008)10:543–550.
- Azari O, Babaei H, Derakhshanfar A, Nematollahi-Mahani SN, Poursahebi R, Moshrefi M. Effects of transplanted mesenchymal stem cells isolated from Wharton's jelly of caprine umbilical cord on cutaneous wound healing; histopathological evaluation. Vet Res Commun (2011) 35:211–22.
- Kim JW, Lee JH, Lyoo YS, Jung DI, Park HM. The effects of topical mesenchymal stem cell transplantation in canine experimental cutaneous wounds. Vet Dermatol (2013) 24:242–e53.
- Maxson S, Lopez EA, Yoo D, Danilkovitch-Miagkova A, LeRoux MA. Role of mesenchymal stem cells in wound repair. Stem Cells Transl Med (2012) 1:142–9.
- Monteiro SO, Lepage OM, Theoret CL. Effects of platelet-rich plasma on the repair of wounds on the distal aspect of the forelimb in horses. Am J Vet Res (2009) 70:277–82.
- Theoret CL, Wilmink JM. Aberrant wound healing in the horse: naturally occurring conditions reminiscent of those observed in man. Wound Repair Regen (2013) 21:365–71.
- Carter-Arnold JL, Neilsen NL, Amelse LL, Odoi A, Dhar MS. In vitro analysis of equine, bone marrow-derived mesenchymal stem cells demonstrates differences within age- and gender-matched horses. Equine Vet J (2014) 46:589–95.
- Ursini TL, Amelse LL, Elkhenany HA, Odoi A, Carter-Arnold JL, Adair HS. Retrospective analysis of local injection site adverse reactions associated with 230 allogenic administrations of bone marrow-derived mesenchymal stem cells in 164 horses. Equine Vet J (2019) 51:198–205.
- Dahlgren LA, Mohammed HO, Nixon AJ. Temporal expression of growth factors and matrix molecules in healing tendon lesions. J Orthop Res (2005) 23:84–92.
- Audette RV, Lavoie-Lamoureux A, Lavoie J-P, Laverty S. Inflammatory stimuli differentially modulate the transcription of paracrine signaling molecules of equine bone marrow multipotent mesenchymal stromal cells. Osteoarthritis Cartilage (2013) 21:1116–24.
- Bowling FL, King L, Fadavi H, Paterson JA, Preece K, Daniel RW. An assessment of the accuracy and usability of a novel optical wound measurement system. Diabet Med (2009) 26:93–6.
- De Schauwer C, Meyer E, Van de Walle GR, Van Soom A. Markers of stemness in equine mesenchymal stem cells: a plea for uniformitiy. Theriogenology (2011) 75:1431–43.
- Dominici ML, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini FC, Krause DS. Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement. Cytotherapy (2006) 8:315–7.
- Moon MH, Kim SY, Kim YJ, Kim SJ, Lee JB, Bae YC. Human adipose tissue-derived mesenchymal stem cells improve postnatal neovascularization in a mouse model of hindlimb ischemia. Cell Physiol Biochem (2006) 17:279–90.
- Lee EY, Xia Y, Kim WS, Kim MH, Kim TH, Kim KJ. Hypoxia-enhanced wound healing function of adipose-derived stem cells: increase in stem cell proliferation and up-regulation of VEGF and bFGF. Wound Repair Regen (2009) 17:540–7.
- Chen L, Tredget EE, Wu PY, Wu Y. Paracrine factors of mesenchymal stem cells recruit macrophages and endothelial lineage cells and enhance wound healing. PLoS ONE (2008) 3:e1886.
- Liu X, Wang Z, Wang R, Zhao F, Shi P, Jiang Y. Direct comparison of the potency of human mesenchymal stem cells derived from amnion tissue, bone marrow and adipose tissue at inducing dermal fibroblast responses to cutaneous wounds. Int J Mol Med (2013) 31:407–15.
- Kim BC, Kim HT, Park SH, Cha JS, Yufit T, Kim SJ. Fibroblasts from chronic wounds show altered TGF-beta-signaling and decreased TGF-beta Type II receptor expression. J Cell Physiol (2003) 195:331–6.
- Caspersen CJ, Thomas GD, Boseman LA, Beckles GL, Albright AL. Aging, diabetes, and the public health system in the United States. Am J Public Health (2012) 102:1482–97.
- Rumalla VK, Borah GL. Cytokines. Growth factors, and plastic surgery. Plast Reconstr Surg (2001) 108:719–33.
- Asahara T, Masuda H, Takahashi T, Kalka C, Pastore C, Silver M. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res (1999) 85:221–8.
- Wilmink JM, van Herten J, van Weeren PR, Barneveld A. Retrospective study of primary intention healing and sequestrum formation in horses compared to ponies under clinical circumstances. Equine Vet J (2002) 34:270–3.
- Chaudhary C, Garg T. Scaffolds: a novel carrier and potential wound healer. Crit Rev Ther Drug Carrier Syst (2015) 32:277–321.
- Chaudhari AA, Vig K, Baganizi DR, Sahu R, Dixit S, Dennis V. Future prospects for scaffolding methods and biomaterials in skin tissue engineering: a review. Int J Mol Sci (2016) 17:1974.
- Rahmani Del Bakhshayesh A, Annabi N, Khalilov R, Akbarzadeh A, Samiei M. Recent advances on biomedical applications of scaffolds in wound healing and dermal tissue engineering. Artif Cells Nanomed Biotechnol (2018) 46:691–705.
- Mund SJ, Kawamura E, Awang-Junaidi AH, Campbell J, Wobeser B, MacPhee DJ. Homing and engraftment of intravenously administered equine cord blood-derived multipotent mesenchymal stromal cells to surgically created cutaneous wound in horses: a pilot project. Cells (2020) 9:1162.
- Textor JA, Clark KC, Walker NJ, Aristizobal FA, Kol A, LeJeune SS. Allogeneic stem cells alter gene expression and improve healing of distal limb wounds in horses. Stem Cells Transl Med (2017) 7:98–108.
- Khashjoori B, Ghamsari SM, Dehghan MM, Farzad Mohajei S, Gholami H, Golshahi H. Evaluation of the effects of adipose derived mesenchymal stem cells cultured on decellularized amniotic membrane in wound healing of distal part of the limbs in horse. Iran J Vet Med (2019) 13:11–25.
Citations
This article has been cited 4 times.- Shi L, Khan MZ, Ullah A, Liang H, Geng M, Akhtar MF, Na J, Han Y, Wang C. Advancements in Stem Cell Applications for Livestock Research: A Review. Vet Sci 2025 Apr 23;12(5).
- Reynolds DE, Vallapureddy P, Morales RT, Oh D, Pan M, Chintapula U, Linardi RL, Gaesser AM, Ortved K, Ko J. Equine mesenchymal stem cell derived extracellular vesicle immunopathology biomarker discovery. J Extracell Biol 2023 Jun;2(6):e89.
- Ribeiro G, Carvalho L, Borges J, Prazeres J. The Best Protocol to Treat Equine Skin Wounds by Second Intention Healing: A Scoping Review of the Literature. Animals (Basel) 2024 May 18;14(10).
- Elkhateeb O, Badawy MEI, Tohamy HG, Abou-Ahmed H, El-Kammar M, Elkhenany H. Curcumin-infused nanostructured lipid carriers: a promising strategy for enhancing skin regeneration and combating microbial infection. BMC Vet Res 2023 Oct 16;19(1):206.
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