The Use of Cord Blood Serum (CBS) in Treating Multiple Conditions, Including Dry Eye Disease and Neuropathy/Neuropathic Eye Pain

Cord Blood Serum (CBS), also reported as Cord blood (CB) or Umbilical Cord Blood (UCB), has been used for the last 10 years as an accepted source of hematopoietic stem cells for the treatment of malignant and non-malignant hematological and immunological diseases (Ref 1-3). 

Cord Blood Serum is thus a life-saving treatment option for certain cancer patients that has now been used in a variety of conditions other including treating rheumatoid arthritis (Ref 2), knee osteoarthritis (Ref 3), potentially blinding ocular diseases as those discussed below (Ref 8-17), multiple sclerosis (Ref 18), autoimmune diseases, such as Rheumatoid arthritis & Lupus, and even glaucoma (Ref 20). 

Cord Blood Serum contains mesenchymal stem cells that can regulate multiple cytokine pathways in response to pro-inflammatory cytokines which can be elevated in a variety of diseases. 

CBS is much easier and less costly to use than bone marrow or even autologous stem cells. Collecting CBS is relatively easy and rapid and has a very low risk of transmission of infections. There is no risk to the donor and a very low risk of graft-vs-host disease with preserved graft-vs-malignancy effects. 

In animal models, CBS has been even shown to repair damaged tissues damaged in heart attacks and strokes (Ref 14,15)
In the eye, CBS has been used to heal the following:
1. Severe epithelial corneal defects
2. Sjogren’s syndrome:
3. Graft Versus Host
4. Recurrent Erosion Syndrome 
5. Persistent corneal 
6. Ocular chemical burns

7. Neurotrophic keratitis
8. Corneal neuropathy
9. Trigeminal neuralgia

CBS eye drops have biologically active components, particularly growth factors which help in wound healing and maintaining normally functioning corneal cells. Cord blood serum is knonw to be more effective than Amniotic Membrane (AM) and/or Amniotic Membrane Extract (AME) in inhibiting inflammatory factors. (Ref 19). 


CBS has been used for years to help with corneal and ocular healing. Research is on going to see if CBS will help regenerate Meibomian glands as well. 


Obtaining reliable sources of CBS has been a challenge and thus limits researchers to continue studying the long term benefits of CBS.
Sandra Lora Cremers, MD, FACS



References:
1. Newcomb JD, Willing AE, Sanberg PR. Umbilical cord blood cells. Methods Mol Biol. 2009;549:119–36.













2. Rocha V, Labopin M, Ruggeri A, et al. Unrelated cord blood transplantation: outcomes after single-unit intrabone injection compared with double-unit intravenous injection in patients with hematological malignancies. Transplantation. 2013;95:1284–91.


























































3. J.A. Brown, V.A. Boussiotis
Umbilical cord blood transplantation: basic biology and clinical challenges to immune reconstitution
Clin Immunol, 127 (2008), pp. 286-297
4. C.G. Brunstein, E.J. Fuchs, S.L. Carter, et al.Alternative donor transplantation after reduced intensity conditioning: results of parallel phase 2 trials using partially HLA-mismatched related bone marrow or unrelated double umbilical cord blood grafts
Blood, 118 (2011), pp. 282-288
5. K.V. Komanduri, L.S. St John, M. de Lima, et al.
Delayed immune reconstitution after cord blood transplantation is characterized by impaired thymopoiesis and late memory T-cell skewing
Blood, 110 (2007), pp. 4543-4551

6.
A. Safdar, G.H. Rodriguez, M.J. De Lima, et al.Infections in 100 cord blood transplantations: spectrum of early and late posttransplant infections in adult and pediatric patients 1996-2005
Medicine (Baltimore), 86 (2007), pp. 324-333

7
A. Stanevsky, G. Goldstein, A. NaglerUmbilical cord blood transplantation: pros, cons and beyond
Blood Rev, 23 (2009), pp. 199-204
8. Körbling M, Robinson S, Estrov Z, et al. Umbilical cord blood-derived cells for tissue repair. Cytotherapy. 2005;7:258–61. 
9. Harris DT. Non-hematological uses of cord blood stem cells. Br J Haematol. 2009;147:177–84.

    Human umbilical cord blood-stem cells direct macrophage polarization and block inflammasome activation to alleviate rheumatoid arthritis. Cell Death & Disease 7. 2016.











11. Yoon KC, Heo H, Im SK, et al. Comparison of autologous serum and umbilical cord serum eye drops for dry eye syndrome. Am J Ophthalmol. 2007;144:86–92. 
12. Versura P, Profazio V, Buzzi M, et al. Efficacy of standardized and quality-controlled cord blood serum eye drop therapy in the healing of severe corneal epithelial damage in dry eye. Cornea. 2013;32:412–8.
13. Yoon KC, Jeong IY, Im SK, et al. Therapeutic effect of umbilical cord serum eyedrops for the treatment of dry eye associated with graft-versus-host disease. Bone Marrow Transplant. 2007;39:231–5.
14. Yoon KC, Im SK, Park YG, et al. Application of umbilical cord blood serum eyedrops for the treatment of dry eye syndrome. Cornea. 2006;25:268–72. 
15. Yoon KC, Choi W, You IC, Choi J. Application of umbilical cord serum eyedrops for recurrent corneal erosions. Cornea. 2011;30:744–8. 
16. Sharma N, Goel M, Velpandian T, et al. Evaluation of umbilical cord serum therapy in acute ocular chemical burns. Invest Ophthalmol Vis Sci. 2011;52:1087–92. 
17. Yoon KC, You IC, Im SK, et al. Application of umbilical cord serum eyedrops for the treatment of neurotrophic keratitis. Ophthalmology. 2007;114:1637–42


18. Meng M, Liu Y, Wang W, Wei C, Liu F, Du Z, Xie Y, Tang W, Hou Z, Li Q. Umbilical cord mesenchymal stem cell transplantation in the treatment of multiple sclerosis.
Am J Transl Res. 2018 Jan 15;10(1):212-223.

19. Tighe S, Moein HR, Chua L, Cheng A, Hamrah P, Tseng SC. Topical cryopreserved amniotic membrane and umbilical cord eye drops
promote re-epithelialization in a murine corneal abrasion model.  Invest
Ophthalmol Vis Sci. 2017;58(3):1586–1593.
25. Rennie K, Gruslin A, Hengstschläger M, et al.  


































20. 
More References: 


Topical Treatment with Cord Blood Serum in Glaucoma Patients: A Preliminary Report

Emilio Campos,1 Piera Versura,1 Giuseppe Giannaccare,1 Adriana Terzi,2Silvia Bisti,3 Stefano Di Marco,3 and Marina Buzzi2
1Ophthalmology Unit, DIMES, Alma Mater Studiorum, University of Bologna, Bologna, Italy
2Emilia Romagna Cord Blood Bank-Transfusion Service, S.Orsola-Malpighi Teaching Hospital, Bologna, Italy
3Vision Lab, DISCAB, University of L’Aquila, L’Aquila, Italy
Correspondence should be addressed to Piera Versura; piera.versura@unibo.it
Received 18 April 2018; Accepted 14 June 2018; Published 25 July 2018
Academic Editor: Nicola Rosa
Copyright © 2018 Emilio Campos et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Abstract

Purpose. To report data which happened to be observed in two glaucoma patients treated with Cord Blood Serum (CBS) eye drops. Design. A case report and retrospective data analysis. Methods. CBS topical eye drops, characterized in advance for growth factors (GFs) content, were administered for two months with the aim to relieve their subjective symptoms, in two patients who had referred ocular surface discomfort, although in absence of any sign of keratopathy. As patients were also affected by advanced glaucoma at risk of vision loss and under treatment with hypotensive drugs, they had been also monitored over the same period with IOP controls and visual field tests in our unit. Results. During subsequent visits, data from Mean Deviation and Pattern Standard Deviation in the visual fields were retrospectively collected and compared with before and after treatment with CBS, and an amelioration was observed. Conclusions. CBS contains a combination of GFs, which potentially exert a neuroprotective action and elect CBS as an interesting natural source to be delivered in neurodegenerative ocular disorders. The incidentally observed amelioration in these two patients deserves further investigation in this respect.





























Brunstein CG. Umbilical cord blood transplantation for the treatment of hematologic malignancies. Cancer Control. 2011;18:222–36



3. Rocha V, Labopin M, Ruggeri A, et al. Unrelated cord blood transplantation: outcomes after single-unit intrabone injection compared with double-unit intravenous injection in patients with hematological malignancies. Transplantation. 2013;95:1284–91. [PubMed] []
4. Körbling M, Robinson S, Estrov Z, et al. Umbilical cord blood-derived cells for tissue repair. Cytotherapy. 2005;7:258–61. [PubMed] []
5. Harris DT. Non-hematological uses of cord blood stem cells. Br J Haematol. 2009;147:177–84.[PubMed] []
6. Yoon KC, Heo H, Im SK, et al. Comparison of autologous serum and umbilical cord serum eye drops for dry eye syndrome. Am J Ophthalmol. 2007;144:86–92. [PubMed] []
7. Versura P, Profazio V, Buzzi M, et al. Efficacy of standardized and quality-controlled cord blood serum eye drop therapy in the healing of severe corneal epithelial damage in dry eye. Cornea. 2013;32:412–8.[PubMed] []
8. Yoon KC, Jeong IY, Im SK, et al. Therapeutic effect of umbilical cord serum eyedrops for the treatment of dry eye associated with graft-versus-host disease. Bone Marrow Transplant. 2007;39:231–5. [PubMed] []
9. Yoon KC, Im SK, Park YG, et al. Application of umbilical cord blood serum eyedrops for the treatment of dry eye syndrome. Cornea. 2006;25:268–72. [PubMed] []
10. Yoon KC, Choi W, You IC, Choi J. Application of umbilical cord serum eyedrops for recurrent corneal erosions. Cornea. 2011;30:744–8. [PubMed] []
11. Sharma N, Goel M, Velpandian T, et al. Evaluation of umbilical cord serum therapy in acute ocular chemical burns. Invest Ophthalmol Vis Sci. 2011;52:1087–92. [PubMed] []
12. Yoon KC, You IC, Im SK, et al. Application of umbilical cord serum eyedrops for the treatment of neurotrophic keratitis. Ophthalmology. 2007;114:1637–42. [PubMed] []
13. Lou-Bonafonte JM, Bonafonte-Marquez E, Bonafonte-Royo S, Martinez-Carpio A. Posology, efficacy, and safety of epidermal growth factor eye drops in 305 patients: logistic regression and group-wise odds of published data. J Ocul Pharmacol Ther. 2012;28(5):467–72. [PubMed] []
14. Shen EP, Hu FR, Lo SC, et al. Comparison of corneal epitheliotrophic capacity among different human blood–derived preparations. Cornea. 2011;30:208–14. [PubMed] []
15. Shlebak AA, Roberts IA, Stevens TA, et al. The impact of antenatal and perinatal variables on cord blood haemopoietic stem/progenitor cell yield available for transplantation. Br J Haematol. 1998;103:1167–71. 


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hUCB-MSCs rarely cause an immune reaction, have high cell activity, are not affected by the patient’s age, and can be extracted at
any time in any quantity [9]. Park et al. reported the results after a seven-year follow-up of repaired cartilage defects




Ref 19:

Human umbilical cord blood-stem cells direct macrophage polarization and block inflammasome activation to alleviate rheumatoid arthritis

Cell Death & Disease 7, e2524 (2016) | Download Citation
Edited by H-U Simon

Abstract

Rheumatoid arthritis (RA) is a long-lasting intractable autoimmune disorder, which has become a substantial public health problem. Despite widespread use of biologic drugs, there have been uncertainties in efficacy and long-term safety. Mesenchymal stem cells (MSCs) have been suggested as a promising alternative for the treatment of RA because of their immunomodulatory properties. However, the precise mechanisms of MSCs on RA-related immune cells are not fully elucidated. The aim of this study was to investigate the therapeutic potential of human umbilical cord blood-derived MSCs (hUCB-MSCs) as a new therapeutic strategy for patients with RA and to explore the mechanisms underlying hUCB-MSC-mediated immunomodulation. Mice with collagen-induced arthritis (CIA) were administered with hUCB-MSCs after the onset of disease, and therapeutic efficacy was assessed. Systemic delivery of hUCB-MSCs significantly ameliorated the severity of CIA to a similar extent observed in the etanercept-treated group. hUCB-MSCs exerted this therapeutic effect by regulating macrophage function. To verify the regulatory effects of hUCB-MSCs on macrophages, macrophages were co-cultured with hUCB-MSCs. The tumor necrosis factor (TNF)-α-mediated activation of cyclooxygenase-2 and TNF-stimulated gene/protein 6 in hUCB-MSCs polarized naive macrophages toward an M2 phenotype. In addition, hUCB-MSCs down-regulated the activation of nucleotide-binding domain and leucine-rich repeat pyrin 3 inflammasome via a paracrine loop of interleukin-1β signaling. These immune-balancing effects of hUCB-MSCs were reproducible in co-culture experiments using peripheral blood mononuclear cells from patients with active RA. hUCB-MSCs can simultaneously regulate multiple cytokine pathways in response to pro-inflammatory cytokines elevated in RA microenvironment, suggesting that treatment with hUCB-MSCs could be an attractive candidate for patients with treatment-refractory RA.

























































. 2014 Jan; 12(Suppl 1): s44–s50.






































Cord blood serum-based eye drops: the impact of donor haematological and obstetric factors on the variability of epidermal growth factor levels





Piera Versura,1 Marina Buzzi,2 Giuseppe Giannaccare,1 Marco Grillini,1 Adriana Terzi,2 Pasqualepaolo Pagliaro,2 andEmilio C. Campos1







Author information ► Article notes ► Copyright and License information ► Disclaimer





















Introduction



Cord blood (CB) is now a recognised and accepted source of haematopoietic stem cells for the treatment of malignant and non-malignant haematological and immunological diseases and a significant increase in its use was recorded over the last decade. The advantages of CB, as compared with bone marrow or mobilised peripheral blood progenitor cells, include ease of collection, rapid availability, low risk of transmission of infections, absence of donor risk and the relatively lower risk of graft-vs-host disease with preserved graft-vs-malignancy effects. However, the relatively small volume and low number of haematopoietic stem cells (HSC) present in a single CB unit have limited the use of CB transplants in adults, unless an intra-bone protocol or double cord transplant is carried out.



Several studies have been performed in animal models suggesting that CB can repair tissues other than blood, in diseases ranging from heart attacks to strokes, but these findings are controversial due to poor repeatability of experiments and it is also not clear how CB may produce such effects.



The pluripotent nature of CB as a biological tear substitute was evidenced in the healing of severe epithelial corneal defects. Several authors have demonstrated the safety and efficacy of CB serum (CBS), topically applied as eye drops, in the treatment of severe dry eye with or without Sjogren’s syndrome,, ocular graft-vs-host disease, persistent corneal epithelial defects, recurrent corneal erosions, chemical burns and neurotrophic keratitis. The rationale for the use of CBS eye drops is their content of biologically active components and in particular of growth factors essential in corneal homeostasis and wound healing, by regulating cellular proliferation, differentiation and migration. Epidermal growth factor (EGF), in particular, plays a central role in corneal wound healing. In vitro experiments have shown the corneal epitheliotropic capacity of different blood-derived preparations and in a previous study we demonstrated the healing efficacy of CBS eyedrops prepared according to a standardised protocol focused on obtaining a daily supply of 0.15 ng/mL EGF, similar to the physiological human tear content. A certain variability in EGF levels in CBS may be present and a preliminary dosage of EGF content in CB units from all donors is performed to pursue the aim of a controlled amount of EGF in the final eye drops. Assaying EGF levels is a time-consuming and expensive procedure and a method for pre-selecting CB units with possibly higher EGF levels, before dosage, is desirable.



Obstetric factors, including gestational age, parity of the mother, sex and birth weight of the neonate, weight of the placenta, duration of labour, and the mode of delivery are known to influence the stem cell content of CB,. It has also been shown that haematological characteristics such as the number of CD34+ stem cells is increased in CB units from Caesarean sections compared to vaginal deliveries,.



The purpose of the present study was to analyse the influence of obstetric and haematological characteristics on the variability of EGF concentration in CB units. This information could aid successful pre-selection of CB units containing higher levels of EGF and, thereby, reduce the number of CB units that need to assayed.













Materials and methods







Data analysis



This is a retrospective study with data from CBS samples obtained from the Emilia Romagna Cord Blood Bank records between 2010 and 2012. Data were collected from 135 donors (median age: 33.1±4.7 years; range, 18 to 42 years).



The following obstetric factors were considered: parity and gestational age of the mother, sex, birth weight and Apgar score of the neonate, placental weight, duration of labour and the mode of delivery (physiological childbirth with vaginal delivery or primary planned Caesarean delivery prior to labour initiation).









Collection method



All CB units were collected after a donor selection questionnaire based on international criteria for CB banking. All steps from the recruitment to the processing and registration of CB were performed according to standard operating procedures and guidelines edited by the Foundation for the Accreditation of Hematopoietic Cellular Therapy (FACT). CB was collected when the placenta was still in utero and the umbilical vein was punctured with a sterile system. The blood was collected under gravitational force into a bag containing 20 mL citrate-phosphate-dextrose (CPD) (Fresenius-Kabi AG, Bad Homburg, Germany). Moreover blood samples were collected from ex utero placental vessels with a sterile syringe and transferred into Vacutest tubes (Kima srl, Padua, Italy) without any anticoagulant. CB units and the related samples were sent for further steps to the processing facility laboratory.









Assessment of cord blood units



The CB units were processed within 48 hours according to CB bank procedures. The presence of markers of maternal infections was determined.









Serum samples



Blood from the ex utero placenta was centrifuged at 3,380 g for 10 minutes and the serum samples obtained were transferred into sterile tubes under a laminar flow hood and stored at −80 °C.









Haematological characteristics



The numbers of total nucleated cells before (TNCb) and after volume reduction (TNCa) were counted with an autoanalyser (Act5/5dif, Beckman Coulter, Cassina de’ Pecchi, Milan, Italy). Erythroblasts were not excluded from the count as it had been estimated that they accounted for less than 3% of the total cells. The number of CD34+ cells was measured by flow cytometry with a single-platform technique (Stem Kit, Beckman Coulter Immunotech, Marseille, France).









Epidermal growth factor dosage



The concentration of EGF in CB units was determined using a Quantikine Human EGF Immunoassay kit (R&D Systems, Inc. Minneapolis, MN, USA). The procedure was carried out according to the manufacturer’s instructions (website: http://www.rndsystems.com/pdf/deg00.pdf) and the absorbance was read at 450 nm using a spectrophotometer (BioRad 680, Hercules, CA, USA).









Statistical analysis



The patients’ characteristics and haematopoietic variables are expressed as mean and standard deviation (SD). Categorical variables (such as sex and parity) were tested by a chi-square test, and continuous variables (such as haematopoietic variables) by the t test. A P value of less than 0.05 was considered to be statistically significant. The statistical analysis was performed with SPSS software, version 14.0 (SPSS Inc., Chicago, IL, USA) and MedCalc 5.0 (MedCalc Software, Ostend, Belgium). Descriptive statistics for tests and variables analysed in subjects are reported as the mean+ SD. Pearson’s (r) or Spearman’s (ρ) correlation coefficients were applied when appropriate; correlations were considered statistically significant for P values <0.05.















Results



The 135 CB units were divided into two groups on the basis of type of delivery: 100 CBS units were derived from physiological vaginal deliveries (group 1) while 35 were derived from planned Caesarean sections (group 2). Maternal (donor) and neonatal characteristics for each group are summarised in Table I. The distribution of mothers’ age, gestational age, child parity and neonatal sex was comparable between the two groups. Similarly, no significant differences were found in neonatal weight and Apgar scores I and II between the two groups.







Table I





Characteristics of the donors (mothers) and neonates.











Characteristics
Group 1: PC
Group 2: CS
P value















Donor
Number
100
35
not done












Maternal age
32.7±4.6 [18–41]
34.6±4.6 [26–42]
0.04












Gestational age (days)
280±7
276±8
0.003












Labour duration (h)
7.62±5.26
0
not applicable












Parity *




 P1
72
22
not done




 P>1
28
13













Newborn
Sex




 Female
46
18
not done




 Male
54
17













Weight (g)
3,599±416
3,556±370





 Female
3,623±412
3,550±315
not significant




 Male
3,574±427
3,561±422













Apgar




 Apgar I
9.13±0.71 [9 (5–10)]
8.97±0.89 [9 (6–10)]
not significant




 Apgar II
9.85±0.42 [10 (8–10)]
9.67±0.58 [10 (8–10)]














Demographic characteristics of the study population (total subjects =135), divided according to the two methods of delivery: group 1: PC = physiological childbirth, vaginal delivery; and group 2: CS = Caesarean section.





*P1 tot = 94 (nulliparous); P>1 tot =41 (primiparous and multiparous).







Haematological variables in the CB were analysed within the 48 hours following collection of the blood. No statistically significant difference was found in the volume of CB collected after the two ways of delivery, nor was the TNC count different between the two groups. Higher numbers of CD34+ cells, TNCb and platelets were found in CB derived from physiological deliveries than from deliveries by Caesarean section, with the difference being statistically significant (P<0.05).



EGF content was statistically significant different between the two groups, with the higher content found in CB derived from physiological delivery (Table II). A certain variability in EGF content was found among donors (Figure 1), with the range being 200–2,000 pg/mL in CB from Caesarean section deliveries and 300–3,000 pg/mL in CB from physiological deliveries.






















An external file that holds a picture, illustration, etc.
Object name is BLT-12-s44-g001.jpg








EGF concentration in CBS obtained after different modes of delivery.



A great variability in EGF content was found among the samples obtained after both vaginal delivery and Caesarean section. A statistically significant difference was demonstrated in EGF content between the two groups, with the higher content found in CBS from vaginal deliveries.













Table II





Maternal and neonatal haematological variables (means ± SD).










Variable
Group 1: PC
Group 2: CS
P value






Volume/CB unit*
108.7±37.76
112.07±30.56
NS




TNCb (×103/μL)
15.73±3.97
13.75±3.63
0.01




TNCa (×106)/CB unit
1,742±516
1,598±360
NS




TNCa (×106)/mL
21.72±14.6
16.98±11.88
NS




CD34+ (×106)/CB unit
4.94±3.3
3.96±1.47
0.02




CD34+ (×106)/mL
0.06±0.05
0.05±0.03
0.03




Platelets (×103/μL)
267.45±58
254.2±52.61
NS




EGF (pg/mL)
1,386±580
1,106±391
0.002













Haematological parameters of the study population divided according to the two methods of delivery: group 1: PC: physiological childbirth, vaginal delivery; and group 2: CS: Caesarean section; CB: cord blood; TNCb: total nucleated cells before volume reduction; TNCa: total nucleated cells after volume reduction; EGF: epidermal growth factor;





*excluding CPD anticoagulant; NS: not significant.







Significant differences (Student’s t test) and correlations (Pearson’s r and Spearman’s ρ) were evaluated between EGF content and patients’ characteristics and between EGF content and haematological parameters.



There was a significant inverse correlation between EGF levels and maternal age (r=−0.25, P=0.005), with a positive predictive value of 91 for age <30 years and of 100 for age <26 years as thresholds for selecting a CB unit with an EGF content higher than 1,000 pg/mL.



A significant inverse correlation was also found between maternal age and both CD34+ cell content/mL (r=−0.36, P<0.0001) and TNC/mL (r=−0.31; P=0.0004).



No correlation was found between EGF content and either gestational age or placental weight.



No differences were found between the groups regarding the neonates’ sex (males vs females), Apgar scores (Apagar I vs Apagar II) and parity (primiparae vs pluriparae).



A direct correlation between EGF levels and CD34+ cells/mL was found when analysing all the CB units included in the study (Figure 2A). A threshold CD34+ cell count of >0.05×106/mL was used in order to identify CB units with an EGF content higher than 1,000 pg/mL (positive predictive value=80). When the data were split into the two groups, the strength of correlation increased in the physiological childbirth group (Figure 2B) while the correlation in the group of CB units from Caesarean section delivery was not statistically significant (Figure 2C).






















An external file that holds a picture, illustration, etc.
Object name is BLT-12-s44-g002.jpg









Correlation between EGF and CD34+ cell content in CBS samples.



A statistically significant correlation was found between the two parameters when all the samples were analysed together (A). When analysing the correlation in samples divided according the method of delivery of the neonate, significance increased in the vaginal delivery group (B) but decreased in the Caesarean section group (C).









EGF content was strongly correlated with the duration of labour in the physiological vaginal delivery group (Figure 3A) as well as with neonatal weight (Figure 3B).






















An external file that holds a picture, illustration, etc.
Object name is BLT-12-s44-g003.jpg









Correlation between EGF content in CBS and duration of labour in hours (A) and between EGF content in CBS and neonatal weight in grams (B).



A strong, statistically significant correlation was found in both cases.









EGF content was also significantly correlated with some haematological parameters such as TNCb/μL (r=0.29, Figure 4A), TNCa/μL (r=0.25, Figure 4B) and platelets/μL (r=0.24, Figure 4C).






















An external file that holds a picture, illustration, etc.
Object name is BLT-12-s44-g004.jpg









Correlation between EGF content in CBS and some haematological parameters: total nucleated cells before volume reduction/μL CB sample (TNCb) (A), total nucleated cells after volume reduction/μL CB sample (TNCa) (B), platelets/mL CB sample (C).



A statistically significant correlation was found in all cases.



















Discussion



Blood derivatives are being currently used as therapeutic agents for several ocular surface diseases and are generally prepared upon request from peripheral blood samples from each single patient. Heterologous blood sources have been recently proposed, with the advantage that they can be routinely obtained as quality/microbiologically-controlled products from blood banks, where a supply may be kept and stored in advance to be readily dispensed.



In a previous study we demonstrated the healing efficacy of CBS-based eye drops obtained from pooled CB units assayed for EGF content, a biologically variable parameter that has to be estimated during the standardised preparation process.



The role of mode of delivery and the effect of foetal distress on the levels of various growth factors and cytokines in CB have been investigated in the past, to understand whether these factors may have an impact on developmental and functional changes in the neonate. Neural growth factor level was found to be significantly lower in preterm infants than in term ones and it was postulated that this growth factor has a role in brain development in later life. In fact, reduced levels of neural growth factor in cord blood may affect foetal growth in preterm deliveries and have implications for neurodevelopmental disorders. Transforming growth factor (TGF)-α and TGF-β1 contribute substantially to normal pregnancy outcomes; higher cord blood TGF-β1 concentrations could be due to increased shear stress in vaginal deliveries, but further studies are needed to evaluate the immunomodulatory role of the labour-associated increase in TGF-β1 in CB.



EGF is detectable in the CB at 23 weeks gestation and its levels rise gradually with increasing gestational age, suggesting that EGF may play a role in foetal growth,.



In the present study a strong relationship was demonstrated between EGF levels and neonatal weight, further suggesting the role of EGF in neonatal development. EGF levels were also strongly related to haematopoietic CD34+ stem cell content and TNC count, and these relationships were statistically significant in CB derived from vaginal deliveries.



We also found that vaginal delivery and labour duration strongly influence EGF content in CB, in agreement with what was reported for haematopoietic CD34+ stem cell content. Our data demonstrated that labour-related foetal distress increases the levels of EGF in CB, in particular when labour lasted longer than 6 hours.



Maternal age also influenced EGF levels: CB from mothers aged less than 26 years contained the highest EGF levels, in agreement with previous findings on the relationship between maternal age and stem cell content.



This study was mainly focused on a search for obstetric and haematological parameters related to both the mother and child that could have an impact on EGF levels in CB, with the goal of pre-selecting CB units appropriate for the preparation of CBS-based eye drops prior to laboratory dosage of EGF. A proper supply of EGF is fundamental for corneal epithelial cell growth in wound healing as shown in in vitroexperiments: EGF enhanced corneal epithelial cell growth in a dose-dependent manner at concentrations from 0.1 to 5 ng/mL; conversely increasing concentrations of EGF from 5 ng/mL to 10 and 100 ng/mL resulted in the down-regulation of clonal growth.



Following the discovery of this dose-dependent response in vitro, in a previous study by our group we standardised a CBS-based eye drop product containing a supply of 0.15 ng/mL EGF per day, to be administered topically for the treatment of severe permanent corneal epithelial defects. A great variability in growth factor content had already been demonstrated; indeed, in order to meet the requirements regarding EGF content, a preliminary phase of assaying and pre-selection of CB units had to be performed so that the final pool of sera included only those containing appropriate EGF levels and excluded those containing too little EGF. This is both time-consuming and expensive and so we investigated alternative ways of making a preliminary selection of CB units.



In the present study we demonstrated that many different obstetric factors must be taken into account when processing CB units for preparation of this particular type of blood component for non-transplantation use. In particular, efforts should be made to select CB units from young mothers, after a labour that lasted longer than 6 hours, and with a haematopoietic CD34+ stem cell content higher than 0.05×106/mL. By applying these three criteria as a whole, we could have been able to pre-select 37 out of 135 CB units, possibly avoiding EGF dosage.













Acknowledgements





The authors thank Mrs Chiara Coslovi for her excellent technical support. The work was supported in part by a grant from the Fondazione Cassa di Risparmio in Bologna to Prof EC Campos and by a grant from the Region of Emilia Romagna to the Cord Blood Bank.















Footnotes









The Authors declare no conflict of interest.



















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Other uses of CBS:



 2019 Jul 22. doi: 10.1111/jocd.13063. [Epub ahead of print]



The effect of human umbilical cord blood-derived mesenchymal stem cell media containing serum on recovery after laser treatment: A double-blinded, randomized, split-face controlled study.



Kim J1,2, Kim B1, Kim S1, Lee YI1,2, Kim J1, Lee JH1,2.





Author information






1


Department of Dermatology and Cutaneous Biology Research Institute, Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.


2


Scar Laser and Plastic Surgery Center, Yonsei Cancer Hospital, Seoul, Korea.













Abstract





BACKGROUND:



Ablative CO2 fractional laser (AFL) is a common cosmetic procedure to improve skin laxity. However, due to prolonged downtime and the risk of postinflammatory hyperpigmentation, laser-assisted delivery of active ingredients as post-laser treatment has gained interest in past years. Among various active ingredients, human umbilical cord blood-derived mesenchymal stem cells (hUCBMSCs) can be a promising agent promoting skin regeneration.



AIMS:



We evaluated the efficacy and safety of a human cord blood cell-conditioned media containing serum and cream on patients who underwent AFL treatment. A randomized, investigator-blinded, prospective, split-face comparison study was conducted.



MATERIALS AND METHODS:



Twenty-three patients who underwent AFL on both cheeks applied a human umbilical cord blood-derived mesenchymal stem cell (hUCBMSC)-conditioned media containing cream with or without stem cell containing serum application. As a primary outcome measure, we evaluated the total area of microcrusts and post-treatment erythema using digital photographs. Additionally, skin biophysical parameters (corneometer, TEWL) and global improvement scores for skin texture were assessed.



RESULTS:



The area of total microcrusts was reduced in the study group which applied both serum and cream. The global improvement score of the post-treatment erythema was significantly reduced. Investigator-assessed global improvement scores were higher in the combination treatment group. Additionally, there was no adverse event, which was associated with the use of either hUCBMSCs containing serum or cream.



CONCLUSION:



The application of human cord blood cell containing serum and cream resulted in accelerated wound healing and reduced post-treatment erythema, which effectively reduced recovery time after ablative laser treatment.





































		
		
			




	




	        			

			
		
	



	


	
 
	

			

			

				

					
					

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