Purpose
To evaluate the platelet capture rate of whole blood fibrin clots and the expression,
secretion, and retention of the growth factors vascular endothelial growth factor
(VEGF), platelet-derived growth factor (PDGF), and basic fibroblast growth factor
(bFGF) from fibrin clots and to determine how these levels may be modulated by allogeneic
adipose-derived stem cells (ASCs).
Methods
Whole blood from 10 human volunteers was transferred to a clotting device and the
platelet capture rate determined. Two experimental conditions and 1 control were evaluated
over 2 weeks in vitro. Clots made from human whole blood without ASCs, clot(−)ASC,
were compared with clots with ASCs incorporated, clot(+)ASC, and a control group of
synthetic polyethylene glycol gels with ASCs incorporated, control(+)ASCs. All conditions
were examined for secretion and retention of VEGF, PDGF, and bFGF via enzyme-linked
immunosorbent assay and immunohistochemistry. The analysis of platelet retention for
clots made with this device was performed.
Results
Enzyme-linked immunosorbent assay analysis showed significantly higher (P < .001) secretion of VEGF in clot(+)ASC compared with clot(−)ASC or control(+)ASC.
In contrast, clot(−)ASC produced soluble PDGF, and the addition of ASCs results in
decreased soluble PDGF with concomitant increases in PDGF immunoreactivity of ASCs.
Soluble bFGF levels were low in clot(−)ASC, and were found to increase at early time
points in clot(+)ASC. Furthermore, bFGF immunoreactivity could be detected in clot(+)ASC,
whereas no bFGF immunoreactivity is present in clot(−)ASC or control(+)ASC. Control(+)ASC
displayed a spike in bFGF secretion at day 0, which may be due to a stress response
elicited by the encapsulation process. Approximately 98% of available platelets in
whole blood were concentrated in the clot on formation.
Conclusions
Fibrin clots made by this method retain high concentrations of platelets, and when
incorporated with ASCs show modulated secretion and immunoreactivity of VEGF, PDGF,
and bFGF.
Clinical Relevance
Whole blood fibrin clots capture platelets and release growth factors, and the addition
of ASCs increases VEGF release for up to 2 weeks after clot formation. This suggests
that whole blood fibrin clots may be a viable scaffold and delivery vehicle for future
stem cell treatments.
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References
- Growth factor delivery methods in the management of sports injuries: The state of play.Br J Sports Med. 2008; 42: 314-320
- Mode of failure for rotator cuff repair with suture anchors identified at revision surgery.J Shoulder Elbow Surg. 2003; 12: 128-133
- Platelet-rich plasma: From basic science to clinical applications.Am J Sports Med. 2009; 37: 2259-2272
Dai R, Wang Z, Samanipour R, Koo K, Kim K. Adipose-derived stem cells for tissue engineering and regenerative medicine applications [published online February 21, 2016]. Stem Cells Int. doi: 10.1155/2016/6737345.
- Platelet-rich plasma for arthroscopic repair of large to massive rotator cuff tears: A randomized, single-blind, parallel-group trial.Am J Sports Med. 2013; 41: 2240-2248
- Adipose-derived regenerative cells promote tendon-bone healing in a rabbit model.Arthroscopy. 2016; 32: 851-859
- Controlled delivery of mesenchymal stem cells and growth factors using a nanofiber scaffold for tendon repair.Acta Biomater. 2013; 9: 6905-6914
- Adipose-derived stem cells improve collagenase-induced tendinopathy in a rat model.Am J Sports Med. 2016; 44: 1983-1989
- Naturally derived and synthetic scaffolds for skeletal muscle reconstruction.Adv Drug Deliv Rev. 2015; 84: 208-221
- A biodegradable fibrin scaffold for mesenchymal stem cell transplantation.Biomaterials. 2003; 24: 2497-2502
- Cytokines in context.J Cell Biol. 1991; 113: 981-986
- Meniscal repair using an exogenous fibrin clot: An experimental study in dogs.J Bone Joint Surg Am. 1998; 70: 1209-1217
- Meniscal repair of degenerative horizontal cleavage tears using fibrin clots: Clinical and arthroscopic outcomes in 10 cases.Orthop J Sports Med. 2014; 2 (2325967114555678)
- Fibrin clot prevents bone tunnel enlargement after ACL reconstruction with allograft.Knee Surg Sports Traumatol Arthrosc. 2017; 25: 1555-1560
- Arthroscopic meniscal repair using an exogenous fibrin clot.Clin Orthop Relat Res. 1990; 252: 64-72
- Polyurethane-covered dacron mesh versus polytetrafluoroethylene DualMesh for intraperitoneal hernia repair in rats.Surg Today. 2002; 32: 884-886
- Arthroscopic inside-out repair of complete radial tears of the meniscus with a fibrin clot.Knee Surg Sports Traumatol Arthrosc. 2013; 21: 2126-2130
- Does fibrin clot really enhance graft healing after double-bundle ACL reconstruction in a caprine model?.Knee Surg Sports Traumatol Arthrosc. 2015; 23: 669-679
- Therapeutic mechanisms of human adipose-derived mesenchymal stem cells in a rat tendon injury model.Am J Sports Med. 2017; 45: 1429-1439
- Runx2-modified adipose-derived stem cells promote tendon graft integration in anterior cruciate ligament reconstruction.Sci Rep. 2016; 6: 19073
- Polymeric growth factor delivery strategies for tissue engineering.Pharm Res. 2003; 20: 1103-1112
- Same or not the same? Comparison of adipose tissue-derived versus bone marrow-derived mesenchymal stem and stromal cells.Stem Cells Dev. 2012; 21: 2724-2752
- Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue.Stem Cells. 2006; 24: 1294-1301
- Adipose-derived stem cells promote angiogenesis and tissue formation for in vivo tissue engineering.Tissue Eng Part A. 2013; 19: 1327-1335
- Adipose-derived stem cells: Implications in tissue regeneration.World J Stem Cells. 2014; 6: 312-321
- Vitronectin-based, biomimetic encapsulating hydrogel scaffolds support adipogenesis of adipose stem cells.Tissue Eng Part A. 2016; 22: 597-609
- Contents and formulations of platelet-rich plasma.Operative Tech Orthop. 2012; 22: 33-42
- Biological strategies to enhance healing of the vascular area of the meniscus.Stem Cells Int. 2012; 2012: 528359
- Rotator cuff repair augmented with endogenous fibrin clot.Arthrosc Tech. 2012; 1: e79-e82
- Partially absorbable meshes for hernia repair offer advantages over nonabsorbable meshes.Am J Surg. 2007; 194: 68-74
- Platelet-rich plasma in bone regeneration: Engineering the delivery for improved clinical efficacy.BioMed Res Int. 2014; 2014: 392398
- Whole blood fibrin clot preserves the viability and bioactivity of human muscle-derived stem cells.ORS Annual Meeting. 2017; (Paper no. 278)
- Heparin-binding domain of fibrin(ogen) binds growth factors and promotes tissue repair when incorporated within a synthetic matrix.Proc Natl Acad Sci USA. 2013; 110: 4563-4568
- Sustained (rh)VEGF 165 release from a sprayed fibrin biomatrix induces angiogenesis, up-regulation of endogenous VEGF-R2, and reduces ischemic flap necrosis.Wound Repair Regen. 2008; 16: 542-550
- FGF-2 but not FGF-1 binds fibrin and supports prolonged endothelial cell growth.J Thromb Haemost. 2003; 1: 1304-1310
- Hypoxia inhibits de novo vascular assembly of adipose-derived stromal/stem cell populations, but promotes growth of preformed vessels.Tissue Eng Part A. 2016; 22: 161-169
- Mechanisms of vasculogenesis in 3D fibrin matrices mediated by the interaction of adipose-derived stem cells and endothelial cells.Angiogenesis. 2014; 17: 921-933
- Concentration of PDGF-AB, BB and TGF-beta1 as valuable human serum parameters in adipose-derived stem cell proliferation.J Nippon Med Sch. 2013; 80: 140-147
- Platelet activation by collagen provides sustained release of anabolic cytokines.Am J Sports Med. 2011; 39: 729-734
- VEGF-A and FGF-2 synergistically promote neoangiogenesis through enhancement of endogenous PDGF-B-PDGFR signaling.J Cell Sci. 2005; 118: 3759-3768
- LHT7, a chemically modified heparin, inhibits multiple stages of angiogenesis by blocking VEGF, FGF2 and PDGF-B signaling pathways.Biomaterials. 2015; 37: 271-278
- The PDGF family: Four gene products form five dimeric isoforms.Cytokine Growth Factor Rev. 2004; 15: 197-204
- Adipose stem cells: Biology and clinical applications for tissue repair and regeneration.Transl Res. 2014; 163: 399-408
- Dose-finding study of fibrin gel-immobilized vascular endothelial growth factor 165 and basic fibroblast growth factor in the arteriovenous loop rat model.Tissue Eng Part A. 2009; 15: 2501-2511
- Vascular endothelial growth factor mediates angiogenic activity during the proliferative phase of wound healing.Am J Pathol. 1998; 152: 1445-1452
- Angiogenic growth factor synergism in a murine tissue engineering model of angiogenesis and adipogenesis.Am J Pathol. 2007; 171: 2048-2057
- Angiogenic synergy of BFGF and VEGF is antagonized by angiopoietin-2 in a modified in vivo matrigel assay.Microvasc Res. 2004; 68: 161-168
- Fibrin-based biomaterials to deliver human growth factors.Thromb Haemost. 2003; 89: 573
- Fibroblast growth factor-2 (FGF-2) induces vascular endothelial growth factor (VEGF) expression in the endothelial cells of forming capillaries: An autocrine mechanism contributing to angiogenesis.J Cell Biol. 1998; 141: 1659-1673
- Differential modulation of cell phenotype by different molecular weight forms of basic fibroblast growth factor: Possible intracellular signaling by the high molecular weight forms.J Cell Biol. 1995; 129: 233-243
Article Info
Publication History
Published online: October 31, 2017
Accepted:
August 3,
2017
Received:
February 9,
2017
Footnotes
The authors report the following potential conflicts of interest or sources of funding: C.S.P. receives $5,000 grant from Pierce Surgical; receives consultancy fees from Arthrex; is a private practitioner at Alta Orthopaedics; and owns stock in Pierce Surgical (less than $10,000, less than 5% of company, no voting or control rights). K.R.S receives grant from U.S. Army Research Office (W911NF-09-0001). Full ICMJE author disclosure forms are available for this article online, as supplementary material.
Identification
Copyright
© 2017 Published by Elsevier on behalf of the Arthroscopy Association of North America
