Decellularized Wharton’s Jelly: Biomaterial Potential for Regenerative Medicine Applications - A Mini-Review

Abstract

The use of Wharton's Jelly (WJ) as a biomaterial is currently undergoing an appearance in the regenerative medicine field. The biomaterials applications focus on the aspects of cellular growth or delivery of proteins capable of stimulating cellular response. However, the basic knowledge about Wharton jelly and decellularization processing technology combined with understanding the physical-chemical properties of this biomaterial is necessary for proper application in regenerative medicine. This mini-review article summarizes information on the composition of WJ, application of drug delivery, in medicine and discusses recent developments with a special focus on its use for regenerative medicine. The most successful and stimulating applications are studies in regenerative medicine and tissue engineering, for wound healing to treat burns, tumor treatment, nanoparticle carriers, and drug delivery systems.

Keywords:
cell seeding; decellularized Wharton's jelly matrix; natural scaffolds; tissue decellularization; Wharton's jelly matrix.

HIGHLIGHTS

• Wharton’s Jelly (WJ) can stimulate cellular response.

• WJ promotes cell proliferation, adhesion, differentiation, and migration.

• WJ 3D printing scaffold can increase the healing tissue potential.

• WJ contains a variety of growth factors that promote tissue regeneration.

INTRODUCTION

Over the past few, sundry regenerative medicine (RM) and nanotechnology approaches have been proposed as potential innovative strategies for many diseases [¹,²]. Biologics presently used in medicinal applications involve platelet-rich plasma, bone marrow aspirate, adipose tissue aspirate, amniotic fluid, amniotic membrane, umbilical cord-derived Wharton's jelly, and cord blood [³,⁴].

Wharton's Jelly (WJ) tissue is a potential biomaterial with increasing use in RM through its decellularization. WG is a fetal tissue composed of mucoid connective tissue around the umbilical cord (UC) composed of collagen, hyaluronic acid (HA) and chondroitin sulfate, and proteoglycans [³]. In this scenario, decellularized WJ constitutes important progress in this research area, as an ideal system to deliver drugs and nanoparticles with growth factors among others and provide a fair environment for cell and tissue regeneration [⁵].

Decellularization is a technique that uses chemical, enzymatic methods, or physical means to extract cellular elements from natural tissues to acquire an acellular extracellular matrix (ECM) scaffold. The product of this process is a three-dimensional (3D) structure of ECM that can be utilized as a biological support (scaffold) for employment in tissue engineering and RM [⁵]. Scaffolds need to be biocompatible, bioabsorbable and with physical and chemical properties that allow cell adhesion, proliferation and differentiation [⁶].

Decellularization processes propose to minimize the tissue potential damage and thus preserve its native mechanical and biologic properties [⁷]. Thus, tissue engineering uses 3D printing with similar properties, different methods, and materials that have remarkable potential applied in tissue implantation. This article intends to resume information on the composition of WJ, its application in medicine, in drug delivery and to debate up to date with emphasis on its use for regenerative medicine.

Characteristic and preparation of Wharton’s jelly

1. Structure

WJ is composed of a layer of mucoid connective gelatinous ECM of the UC. It is composed of a layer of simple amniotic epithelium and matrix elements such as collagen (types I, III, IV, and V), HA, and various sulfated glycosaminoglycans that surround the UC vessels. The structural integrity of these components does not alter after the current decellularization process (Figure 1) [⁸,⁹].

Figure 1
The structural integrity of Wharton's Jelly before and after the decellularization process.

2. Properties

Wharton jelly mesenchymal stem cells have several functions in tissue reconstruction in ophthalmologic surgery and wound treatment. Biomechanically, its resistance and elasticity provide biochemical structure and protection to the epithelial cells. Biologically, promote osteogenic and chondrogenic cells' adhesion, differentiation, and migration, and stimulates extracellular matrix production in vivo. Moreover, it has the properties of pro-inflammatory and anti-inflammatory cytokines with are essential for proper tissue healing [¹⁰]. Recent studies show that WJ, expressed growth factor-1 (IGF-1), but also platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), and TGF-α, (transforming growth factor - α [¹¹].

With its immunomodulatory effect, GW can stimulate different tissues. IGF-1 induces osteogenic and chondrogenic differentiation of mesenchymal stem cells and improves extracellular matrix propagation [³]. Transforming growth factor (GF), through its receptor, promotes proliferation and survival of osteoprogenitors and plays an anabolic role in bone metabolism [⁴]. PDGF exhibits chemotactic properties toward human osteoblasts, and its downregulation is related to cartilage deterioration [¹²]. VEGF, a signal protein that induces angiogenesis, is included in tissue remodeling and bone formation [¹¹].

Furthermore, several pro-inflammatory cytokines associated with healing in wounds were identified in the WG. The intercellular adhesion molecule-1 (ICAM-1) promotes leukocyte accumulation in the wound site and has immunosuppressive effects on dendritic cells and T cells, which may aid in the treatment of graft versus host diseases [¹³,¹⁴]. The expression of monocyte chemotactic protein1 (aka CCL2), a cytokine, induces wound healing as well, including in diabetic wounds as demonstrated in a previous study [¹⁵].

Umbilical cord WJ seems to be an optimal biomaterials source for cell culture systems and in the application as three-dimensional scaffolding. Since studies validated that it is a biocompatible matrix that permits cellular attachment, development, and proliferation with satisfactory properties in vitro and in vivo. Furthermore, it can be painlessly harvested in abundance in several births around the world without generating morbidity to the donor [¹⁶,¹⁷].

3. Preparation, sterilization, and preservation

a. Preparation

The method of umbilical cord (UC) preparation was as previously described by Bartolucci. In brief, the donor patience of the placenta is selected by serological tests to exclude human immunodeficiency virus (HIV), hepatitis B, hepatitis C, or syphilis). The placenta is broadly flushed in sterile PBS containing 200 U/mL penicillin and 200 μg/mL streptomycin. Then the WJ was sectioned into small-scale parts (1-2 mm), implanted in 100-mm culture plates. This method is used in various studies with few changes [¹⁸].

b. Sterilization

The UC is collected under sterile conditions. Normally, to maintain the UC and WJ under sterile conditions are used antibiotics and antimycotics cover gram-positive and negative bacteria and fungi to prevent microbial proliferation from contamination during processing. Additionally, it is pointed out immediately refrigeration at 4 ^oC after the sterilization process. It is also reported the initiation of the decellularization process within 72h after UC collection [¹⁹].

c. Preservation

Previous research showed that the protocols for cryopreservation are the most common method of preservation of the decellularized umbilical cord segments. Thus, the cryopreservation substance consists of saline without cryoprotectants. It is utilized in > 20 volume excess to the volume of fresh material at a cooling rate of 1 °C/min, and before it is stored at - 20°С [²⁰,²¹]. Most effective protocols utilize CryoStor CS10 (which contains 10% DMSO), freezing to - 80 °С, storage in liquid nitrogen for 1 month and rapid thawing at 37 °C. Also, decellularized WJ can be preserved using a lyophilized technique (freeze-drying) [¹²].

4. Clinical Applications of WJ in surgical treatment

Wharton's jelly is a connective gelatinous umbilical cord tissue. Tissue engineering aims to use the principles and technologies for obtaining scaffolds useful for regeneration medicine applications. Wharton's jelly in medical applications is extensively used because it has good biocompatibility and low cost. Even after the decellularization process, the matrix of WJ retains a diversity of bioactive substances like GF, collagen, laminin, fibronectin, and polysaccharide. Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) in situ transplantation promoted functional cardiac recovery after AMI [ ²²,²³].

In the same manner, Hashemi and coauthors 2019 in observed that (WJSCs) seeded on an amniotic membrane could promote a healing effect in diabetic wounds [²⁴]. The application of WJ-MSCs enclosed with PF-127/SAP also promoted cell proliferation, neovascularization, and wound diabetic healing in rats [²⁵].

Wang and coauthors 2013, in a previous study, demonstrated that the WJ-MSCs utilization improved the neurological function in rats with spinal cord injury [²⁶]. Furthermore, a multi-center trial study has shown excellent results concerning the safety and efficacy of transplanted WJ-MSCs in patients with knee osteoarthritis [²⁷]. The advantage of WJ-MSCs for cartilage healing gives indications to be the most promising utilization for this biomaterial, given the similarities between chondrocytes, WJ cells and the cellular matrix of cartilage [²⁸].

In addition, a preliminary in vivo study demonstrated that WJ-MSC can differentiate into neuron-like cells, endothelial-like cells, myocyte-like cells, insulin-producing cells, and hepatocyte-like cells. However, there are still few studies correlating the potential of WJMSCs with mesenchymal stem cells came of other tissue origins [²⁹].

A systematic review compared WJ-MSCs with other sources of MSCs in the treatment of cancer and showed that WJ-MSCs have more antitumorigenic effects. Despite that, there are still no clinical tests using WJ-MSCs for that purpose. Since, albeit 88% of the research showed that the WJ-MSCs allow for reducing cancer cell proliferation, 12% of the papers, which are all in vitro studies, reported contradicting results whereby WJ-MSCs improved cancer spreading [³⁰].

CONCLUSION

Several publications have suggested that the application of WJ accelerates the onset of the cell proliferative in chronic diabetic wounds. Experimental studies using WJ after acute myocardial infarction have described promising results in cardiac function in recent years.

Finally, we note that experimental and clinical trials have intensively explored Wharton's jelly mesenchymal stem cells as a matrix for promising three-dimensional scaffolding migration, proliferation, and cell differentiation in regenerative medicine.

• Funding: This research received no external funding.

Acknowledgments:

We thank the Health Sciences Postgraduate Program at the Pontifical Catholic University of Paraná for encouraging this research.

REFERENCES

• ¹
  Davies JE, Walker JT, Keating A. Concise Review: Wharton's Jelly: The Rich, but Enigmatic, Source of Mesenchymal Stromal Cells. Stem Cells Transl Med. 2017 Jul;6(7):1620-30.
• ²
  Mohammadie ZM, Parivar K, Shahri NM, Fereidoni M, Hayati-Roodbari N. Decellularized Bovine Articular Cartilage Matrix Reinforced by Carboxylated-SWCNT for Tissue Engineering Application. Braz Arch Biol Technol [Internet]. 2018 Jan 8 [cited 2022 Apr 29];60:17160083.
• ³
  Fortier LA, Barker JU, Strauss EJ, McCarrel TM, Cole BJ. The role of growth factors in cartilage repair. Clin Orthop Relat Res. 2011;469:2706-15.
• ⁴
  Zhang X, Tamasi J, Lu X, Zhu J, Chen H, Tian X, et al. Epidermal growth factor receptor plays an anabolic role in bone metabolism in vivo. J Bone Miner Res. 2011;26:1022-34.
• ⁵
  Main BJ, Maffulli N, Valk JA, Rodriguez HC, Gupta M, El-Amin 3 rd SF, et al. Umbilical cord-derived Wharton's jelly for regenerative medicine applications: A systematic review. Pharmaceuticals. 2021;14(11):1-5.
• ⁶
  Braga SF, Trovatti E, de Carvalho RA, de Carvalho AJF, Iemma MR da C, Amaral AC. Bioactive Fibrin Scaffolds for Use in Musculoskeletal Regenerative Medicine. Braz Arch Biol Technol [Internet]. 2020 Aug 10 [cited 2022 Apr 29];63:2020.
• ⁷
  Converse GL, Li D, Buse EE, Hopkins RA, Aljitawi OS. Wharton's Jelly Matrix Decellularization for Tissue Engineering Applications. Methods Mol Biol. 2018;1577:25-33.
• ⁸
  Edgar L, Pu T, Porter B, Aziz JM, La Pointe C, Asthana A, Orlando G. Regenerative medicine, organ bioengineering, and transplantation. Br J Surg. 2020;107(7):793-800.
• ⁹
  Wang LR, Lin YQ, Wang JT, Pan LL, Huang KT, Wan L, et al. Recent advances in the re-engineered liver: de-cellularization and re-cellularization techniques. Cytotherapy. 2015 Aug;17(8):1015-24.
• ¹⁰
  Spiller KL, Nassiri S, Witherel CE, Anfang RR, Ng J, Nakazawa KR, et al. Sequential delivery of immunomodulatory cytokines to facilitate the M1-to-M2 transition of macrophages and enhance vascularization of bone scaffolds. Biomaterials. 2015 Jan;37: 194-207.
• ¹¹
  Wyatt LA, Nwosu LN, Wilson D, Hill R, Spendlove I, Bennett AJ, et al. Molecular expression patterns in the synovium and their association with advanced symptomatic knee osteoarthritis. Osteoarthritis Cartilage. 2019 Apr;27(4):667-75.
• ¹²
  Lind M. Growth factor stimulation of bone healing. Effects on osteoblasts, osteomies, and implants fixation. Acta Orthop Scand Suppl. 1998;283:2-37.
• ¹³
  Nagaoka T, Kaburagi Y, Hamaguchi Y, Hasegawa M, Takehara K, Steeber DA, et al. Delayed wound healing in the absence of intercellular adhesion molecule-1 or L-selectin expression. Am J Pathol. 2000;157:237-47.
• ¹⁴
  Tang B, Li X, Liu Y, Chen X, Li X, Chu Y, et al. The therapeutic effect of ICAM-1-overexpressing mesenchymal stem cells on acute graft-versus-host disease. Cell Physiol Biochem. 2018;46:2624-35.
• ¹⁵
  Wood S, Jayaraman V, Huelsmann EJ, Bonish B, Burgad D, Sivaramakrishnan G, et al. Pro-inflammatory chemokine CCL2 (MCP-1) promotes healing in diabetic wounds by restoring the macrophage response. PLoS One. 2014;9:e91574.
• ¹⁶
  Jadalannagari S, Converse G, McFall C, Buse E, Filla M, Villar MT, et al. Decellularized Wharton's Jelly from human umbilical cord as a novel 3D scaffolding material for tissue engineering applications. PLoS One. 2017;12(2):e0172098.
• ¹⁷
  Liau LL, Ruszymah BHI, Ng MH, Law JX. Characteristics and clinical applications of Wharton's jelly-derived mesenchymal stromal cells. Curr Res Transl Med. 2020;68(1):5-16.
• ¹⁸
  Bartolucci J, Verdugo FJ, González PL, Larrea RE, Abarzua E, Goset C, et al. Safety and Efficacy of the Intravenous Infusion of Umbilical Cord Mesenchymal Stem Cells in Patients With Heart Failure: A Phase 1/2 Randomized Controlled Trial (RIMECARD Trial [Randomized Clinical Trial of Intravenous Infusion Umbilical Cord Mesenchymal Stem Cells on Cardiopathy]). Circ Res. 2017 Oct 27;121(10):1192-1204.
• ¹⁹
  Puzanov MV, Vasilyeva LB, Popova PV, Grineva EN, Dmitrieva RI. New Approach to Cryopreservation of Primary Noncultivated Human Umbilical Vein Endothelium in Biobanking. Biopreserv Biobank. 2018 Apr;16(2):114-119.
• ²⁰
  Tuan-Mu HY, Yu CH, Hu JJ. On the decellularization of fresh or frozen human umbilical arteries: implications for small-diameter tissue engineered vascular grafts. Ann Biomed Eng. 2014 Jun;42(6):1305-18.
• ²¹
  Arutyunyan I, Fatkhudinov T, Sukhikh G. Umbilical cord tissue cryopreservation: a short review. Stem Cell Res Ther. 2018 Sep 15;9(1):236.
• ²²
  Couture M. A Single-center, Retrospective Study of Cryopreserved Umbilical Cord for Wound Healing in Patients Suffering from Chronic Wounds of the Foot and Ankle. Wounds. 2016 Jul;28(7):217-25.
• ²³
  Rabbani S, Soleimani M, Sahebjam M, Imani M, Haeri A, Ghiaseddin A, et al. Simultaneous Delivery of Wharton's Jelly Mesenchymal Stem Cells and Insulin-Like Growth Factor-1 in Acute Myocardial Infarction. Iran J Pharm Res. 2018 Spring;17(2):426-41.
• ²⁴
  Hashemi SS, Mohammadi AA, Kabiri H, Hashempoor MR, Mahmoodi M, Amini M, et al. The healing effect of Wharton's jelly stem cells seeded on biological scaffold in chronic skin ulcers: A randomized clinical trial. J Cosmet Dermatol. 2019 Dec;18(6):1961-7.
• ²⁵
  Jiao Y, Chen X, Niu Y, Huang S, Wang J, Luo M, et al. Wharton's jelly mesenchymal stem cells embedded in PF-127 hydrogel plus sodium ascorbyl phosphate combination promote diabetic wound healing in type 2 diabetic rat. Stem Cell Res Ther. 2021 Oct 30;12(1):559.
• ²⁶
  Wang S, Cheng H, Dai G, Wang X, Hua R, Liu X, et al. Umbilical cord mesenchymal stem cell transplantation significantly improves neurological function in patients with sequelae of traumatic brain injury. Brain Res. 2013 Sep 26;1532:76-84.
• ²⁷
  Gupta A, Maffulli N, Rodriguez HC, Lee CE, Levy HJ, El-Amin 3rd SF. Umbilical cord-derived Wharton's jelly for treatment of knee osteoarthritis: study protocol for a non-randomized, open-label, multi-center trial. J Orthop Surg Res. 2021 Feb 18;16(1):143.
• ²⁸
  Li Z, Bi Y, Wu Q, Chen C, Zhou L, Qi J, et al. A composite scaffold of Wharton’s jelly and chondroitin sulphate loaded with human umbilical cord mesenchymal stem cells repairs articular cartilage defects in rat knee. J Mater Sci Mater Med. 2021 Apr 1;32(4).
• ²⁹
  Rezaeian L, Hosseini SE, Dianatpour M, Edalatmanesh MA, Tanideh N, Mogheiseh A, et al. Intrauterine xenotransplantation of human Wharton jelly-derived mesenchymal stem cells into the liver of rabbit fetuses: a preliminary study for in vivo expression of the human liver genes. Iran J Basic Med Sci 2018;21:89-9
• ³⁰
  Christodoulou I, Goulielmaki M, Devetzi M, Panagiotidis M, Koliakos G, Zoumpourlis V. Mesenchymal stem cells in preclinical cancer cytotherapy: a systematic review. Stem Cell Res Ther 2018;9:336.

Editor-in-Chief: Paulo Vitor Farago

Associate Editor: Paulo Vitor Farago

Publication Dates

• Publication in this collection
  19 Sept 2022
• Date of issue
  2023

History

• Received
  20 May 2022
• Accepted
  29 June 2022