Development of a dual growth factor loaded biodegradable hydrogel and its evaluation on osteoblast differentiation in vitro

2011 ◽  
Vol 1312 ◽  
Author(s):  
Deepti Dyondi ◽  
Thomas J Webster ◽  
Rinti Banerjee
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Osteoblast Differentiation ◽  
Chitosan Nanoparticles ◽  
Marker Genes ◽  
Tissue Engineering Scaffolds ◽  
Biodegradable Hydrogel ◽  
Growth Factor Release ◽  
Factor Release

ABSTRACTHydrogels with their tunable properties are attractive candidates for developing tissue engineering scaffolds for various applications (including bone and cartilage). The current work involved studying the synergistic effect of basic fibroblast growth factor (bFGF) and platelet derived growth factor BB (PDGF-BB) entrapped within injectable porous gels for bone regeneration applications. An in situ gelling system was developed using bacterial polysaccharides gellan and xanthan gum by temperature and ionic gelation with Ca+2. After the initial characterization of the hydrogels, a dual growth factor release system was developed wherein growth factors were encapsulated within chitosan nanoparticles embedded in the gels as well as directly within the gel. The hydrogel structure was characterized by SEM and TEM and in vitro growth factor release studies showed a slow release profile in PBS. Further, human fetal osteoblasts were entrapped within the hydrogel and a 21 day osteoblast differentiation study was conducted. An improvement in osteoblast total protein synthesis and collagen content was observed by day 21 compared to control gels without growth factors. Although further evaluation regarding mechanical properties and expression of osteogenic differentiation marker genes will be necessary, the present study suggests that injectable scaffolds can be used for the delivery of multiple growth promoting agents to support osteoblast differentiation.

Bone-derived growth factor release from poly(α-hydroxy acid) implants in vitro

Biomaterials ◽  
1993 ◽  
Vol 14 (3) ◽  
pp. 177-183 ◽  
Author(s):  
M.C. Meikle ◽  
W.-Y. Mak ◽  
S. Papaioannou ◽  
E.H. Davies ◽  
N. Mordan ◽  
...  
Keyword(s):  
Growth Factor ◽  
Hydroxy Acid ◽  
Growth Factor Release ◽  
Factor Release

scholarly journals Biofunctionalization of porcine-derived collagen matrices with platelet rich fibrin: influence on angiogenesis in vitro and in vivo

2020 ◽  
Vol 24 (10) ◽  
pp. 3425-3436 ◽  
Author(s):  
Sebastian Blatt ◽  
Valentin Burkhardt ◽  
Peer W. Kämmerer ◽  
Andreas M. Pabst ◽  
Keyvan Sagheb ◽  
...  
Keyword(s):  
Growth Factor ◽  
Immunohistochemical Staining ◽  
Collagen Matrices ◽  
Platelet Rich Fibrin ◽  
Angiogenic Potential ◽  
Branching Points ◽  
Growth Factor Release ◽  
Factor Release

Abstract Objectives Porcine-derived collagen matrices (CM) can be used for oral tissue regeneration, but sufficient revascularization is crucial. The aim of this study was to analyze the influence of platelet-rich fibrin (PRF) on angiogenesis of different CM in vitro and in vivo. Materials and methods Three different CM (mucoderm, jason, collprotect) were combined with PRF in a plotting process. Growth factor release (VEGF, TGF-β) was measured in vitro via ELISA quantification after 1,4 and 7 days in comparison to PRF alone. In ovo yolk sac (YSM) and chorion allantois membrane (CAM) model, angiogenic potential were analyzed in vivo with light- and intravital fluorescence microscopy after 24 h, then verified with immunohistochemical staining for CD105 and αSMA. Results Highest growth factor release was seen after 24 h for all three activated membranes in comparison to the native CM (VEGF 24 h: each p < 0.05; TGF-β: each p < 0.001) and the PRF (no significant difference). All activated membranes revealed a significantly increased angiogenic potential in vivo after 24 h (vessels per mm2: each p < 0.05; branching points per mm2: each p < 0.01; vessel density: each p < 0.05) and with immunohistochemical staining for CD105 (each p < 0.01) and αSMA (each p < 0.05). Conclusions PRF improved the angiogenesis of CM in vitro and in vivo. Clinical relevance Bio-functionalization of CM with PRF could easily implemented in the clinical pathway and may lead to advanced soft tissue healing.

Effects of Aspirin on Growth Factor Release From Freshly Isolated Leukocyte-Rich Platelet-Rich Plasma in Healthy Men: A Prospective Fixed-Sequence Controlled Laboratory Study

2019 ◽  
Vol 47 (5) ◽  
pp. 1223-1229 ◽  
Author(s):  
Prathap Jayaram ◽  
Peter Yeh ◽  
Shiv J. Patel ◽  
Racel Cela ◽  
Theodore B. Shybut ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Whole Blood ◽  
Low Dose ◽  
Platelet Rich Plasma ◽  
Healthy Men ◽  
Growth Factor Release ◽  
Tgf Β1 ◽  
Cox 1 ◽  
Factor Release

Background: The benefits of platelet-rich plasma (PRP) are believed to be in part dependent on growth factor release after platelet activation. Platelet activation is complex and involves multiple mechanisms. One important mechanism is driven by cyclooxygenase 1 (COX-1)–mediated conversion of arachidonic acid (AA) to precursor prostaglandins that then mediate proinflammatory responses that trigger growth factor release. Acetylsalicylic acid (ASA; also known as aspirin) is known to irreversibly inhibit COX-1, thereby blocking AA-mediated signaling; however, it is unclear whether ASA use alters growth factor release from freshly isolated PRP. Purpose: To assess the effects of low-dose ASA use on activation of growth factor release from freshly isolated human PRP via AA and thrombin (TBN). Study Design: Controlled laboratory study. Methods: Twelve healthy men underwent blood collection and leukocyte-rich PRP (LR-PRP) preparation through a double-spin protocol to obtain baseline whole blood and PRP counts the same day. PRP was aliquoted into 3 groups: nonactivated, AA activated, and TBN activated. Immediately after activation, the concentrations of transforming growth factor β1 (TGF-β1), vascular endothelial growth factor (VEGF), and platelet-derived growth factor AB (PDGF-AB) were measured using enzyme-linked immunosorbent assays (ELISAs). The same 12 participants were then placed on an 81-mg daily dose of oral ASA for 14 days. Repeat characterization of whole blood and PRP analyses was done on day 14, followed by repeat ELISAs of growth factors under the same nonactivated and activated settings as previously stated. Results: Fourteen days of daily ASA had no effect on the number of platelets and leukocytes measured in whole blood and LR-PRP. Compared with nonactivated LR-PRP, AA- and TBN-mediated activation led to significant release of VEGF and PDGF-AB. In contrast, release of TGF-β1 from LR-PRP was observed only with activation by AA, not with TBN. Consistent with its inhibitory role in AA signaling, ASA significantly inhibited AA-mediated release of all 3 growth factors measured in this study. Although ASA had no effect on TBN-mediated release of VEGF and TGF-β1 from LR-PRP, ASA did partially block TBN-mediated release of PDGF-AB, although the mechanism remains unclear. Conclusion: Daily use of low-dose ASA reduces VEGF, PDGF-AB, and TGF-β1 expression in freshly isolated human LR-PRP when activated with AA. Clinical Relevance: Reduction in growth factor release attributed to daily use of low-dose ASA or other COX inhibitors can be mitigated when PRP samples are activated with TBN. Clinical studies are needed to determine whether activation before PRP injection is needed in all applications where ASA is in use and to what extent ASA may inhibit growth factor release in vivo at the site of injury.

Comparison of Release Profiles of Various Growth Factors from Biodegradable Carriers

1998 ◽  
Vol 530 ◽  
Author(s):  
Y. Tabata ◽  
M. Yamamoto ◽  
Y. Ikada
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
The Body ◽  
Bone Morphogenetic Protein 2 ◽  
Biodegradable Hydrogel ◽  
Tgf Β1

AbstractA biodegradable hydrogel was prepared by glutaraldehyde crosslinking of acidic gelatin with an isoelectric point (IEP) of 5.0 as a carrier to release basic growth factors on the basis of polyion complexation. Basic fibroblast growth factor (bFGF), transforming growth factor β1 (TGF-β1), and bone morphogenetic protein-2 (BMP-2) were sorbed from their aqueous solution into the dried gelatin hydrogels to prepare respective growth factor-incorporating hydrogels. Under an in vitro non-degradation condition, approximately 20 % of incorporated bFGF and TGF-β1 was released from the hydrogels within initial 40 min, followed by no further release, whereas a large initial release of BMP-2 was observed. After subcutaneous implantation of the gelatin hydrogels incorporating 125I-labeled growth factor in the mouse back, the remaining radioactivity was measured to estimate the in vivo release profile of growth factors. Incorporation into gelatin hydrogels enabled bFGF and TGF-β1 to retain in the body for about 15 days and the retention period well correlated with that of the gelatin hydrogel. Taken together, it is likely that the growth factors ionically complexed with acidic gelatin were released in vivo as a result of hydrogel biodegradation. On the contrary, basic BMP-2 did not ionically interact with acidic gelatin, resulting in no sustained released by the present biodegradable carrier system.

scholarly journals Heparin‐regulated growth factor release in vitro and angiogenesis in vivo from hyaluronan hydrogels

2007 ◽  
Vol 21 (5) ◽  
Author(s):  
Daniel B. Pike ◽  
Shenshen Cai ◽  
Kyle R. Pomraning ◽  
Matthew A. Firpo ◽  
Xiao Zheng Shu ◽  
...  
Keyword(s):  
Growth Factor ◽  
Growth Factor Release ◽  
Release In Vitro ◽  
Factor Release

scholarly journals Nanodiamond-based injectable hydrogel for sustained growth factor release: Preparation, characterization and in vitro analysis

2017 ◽  
Vol 58 ◽  
pp. 479-491 ◽  
Author(s):  
Settimio Pacelli ◽  
Francisca Acosta ◽  
Aparna R. Chakravarti ◽  
Saheli G. Samanta ◽  
Jonathan Whitlow ◽  
...  
Keyword(s):  
Growth Factor ◽  
Injectable Hydrogel ◽  
Sustained Growth ◽  
Release Preparation ◽  
Growth Factor Release ◽  
Vitro Analysis ◽  
In Vitro Analysis ◽  
Factor Release

scholarly journals P5: EFFECT OF VITAMIN C AND COLD ON GROWTH FACTOR RELEASE FROM PLATELET-RICH PLASMA

2021 ◽  
Vol 108 (Supplement_1) ◽  
Author(s):  
T Tomouk ◽  
S Talaat ◽  
O Smith ◽  
A Mosahebi
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Vitamin C ◽  
Sustained Release ◽  
Platelet Rich Plasma ◽  
Growth Factor Release ◽  
Cold Pack ◽  
B Vitamin ◽  
Factor Release ◽  
Vit C

Abstract Introduction Platelet-rich plasma (PRP) has gathered great momentum in surgery. The main outcome of the study was to assess whether there was a difference in growth factor release from PRP with addition of vitamin C, cooling, or both vitamin C and cooling. Method Blood samples from 10 volunteers were centrifuged to produce PRP. Activated autologous thrombin was produced from platelet-poor plasma (PPP), which was added to PRP to activate it. PRP from each participant was then divided by activation method into three groups: A) cold pack only; B) vitamin C only; C) cold pack/vitamin C. Growth factor quantification was performed for human VEGF, IGF-1 and PDGF-AB using enzyme-linked immunosorbent assays. Result At 24 hours, VEGF concentration in the cold group was significantly higher cf. the vitamin C group (p=0.02) and the cold/vit C group (p&lt;0.01), whereas IGF concentration in the cold group was significantly higher cf. the vitamin C group (p=0.02) but not the cold/vit C group. At 1 hour and 24 hours, PDGF concentration in the cold group was significantly higher cf. both the vitamin C group (p&lt;0.01) and the cold/vit C group (p=0.02 and p&lt;0.0001 respectively). Conclusion Cooling lead to increased growth factor release at 24 hours, indicating that cooling may lead to sustained release of growth factors. There was a trend for PRP treated with vitamin C to remain in a gel-like consistency rather than clot. This may prove useful in clinical practice where currently activated PRP is constrained to topical application only due to its consistency. Take-home message Cooling may lead to sustained release of growth factors from platelet-rich plasma, whereas the addition of vitamin C may provide opportunity for injectable activated PRP.

scholarly journals Matrix-Bound Growth Factors are Released upon Cartilage Compression by an Aggrecan-Dependent Sodium Flux that is Lost in Osteoarthritis

2021 ◽  
Author(s):  
Stuart J. Keppie ◽  
Jessica C. Mansfield ◽  
Xiaodi Tang ◽  
Christopher J. Philp ◽  
Helen K. Graham ◽  
...  
Keyword(s):  
Growth Factors ◽  
Articular Cartilage ◽  
Growth Factor ◽  
Heparan Sulfate ◽  
Tissue Growth ◽  
Acoustic Microscopy ◽  
Matrix Stiffness ◽  
Sodium Flux ◽  
Growth Factor Release ◽  
Factor Release

AbstractArticular cartilage is a dense extracellular matrix-rich tissue that degrades following chronic mechanical stress, resulting in osteoarthritis (OA). The tissue has low intrinsic repair especially in aged and osteoarthritic joints. Here we describe three pro-regenerative factors; fibroblast growth factor 2 (FGF2), connective tissue growth factor, bound to transforming growth factor-beta (CTGF-TGFβ), and hepatoma-derived growth factor (HDGF), that are rapidly released from the pericellular matrix (PCM) of articular cartilage upon mechanical injury. All three growth factors bound heparan sulfate, and were displaced by exogenous NaCl. We hypothesised that sodium, sequestered within the aggrecan-rich matrix, was freed by injurious compression, thereby enhancing the bioavailability of pericellular growth factors. Indeed, growth factor release was abrogated when cartilage aggrecan was depleted by IL-1 treatment, and in severely damaged human osteoarthritic cartilage. A flux in free matrix sodium upon mechanical compression of cartilage was visualised by 23Na magnetic resonance imaging (MRI) just below the articular surface. This corresponded to a region of reduced tissue stiffness, measured by scanning acoustic microscopy and second harmonic generation microscopy, and where Smad2/3 was phosphorylated upon cyclic compression. Our results describe a novel intrinsic repair mechanism, controlled by matrix stiffness and mediated by the free sodium concentration, in which heparan sulfate-bound growth factors are released from cartilage upon injurious load. They identify aggrecan as a depot for sequestered sodium, explaining why osteoarthritic tissue loses its ability to repair. Treatments that restore matrix sodium to allow appropriate release of growth factors upon load are predicted to enable intrinsic cartilage repair in osteoarthritis.Significance StatementOsteoarthritis is the most prevalent musculoskeletal disease, affecting 250 million people worldwide 1. We identify a novel intrinsic repair response in cartilage, mediated by aggrecan-dependent sodium flux, and dependent upon matrix stiffness, which results in the release of a cocktail of pro-regenerative growth factors after injury. Loss of aggrecan in late-stage osteoarthritis prevents growth factor release and likely contributes to disease progression. Treatments that restore matrix sodium in osteoarthritis may recover the intrinsic repair response to improve disease outcome.

scholarly journals THE G-FORCE CONUNDRUM IN PRF GENERATION- MANAGEMENT OF A PROBLEM HIDDEN IN PLAIN SIGHT

2020 ◽  
Author(s):  
Kidambi Sneha ◽  
Jhansi Rani Ajmera ◽  
Rampalli Viswa Chandra
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Time Period ◽  
Significant Difference ◽  
Platelet Concentration ◽  
Size Growth ◽  
Growth Factor Release ◽  
Optimum Relationship ◽  
Centrifugation Protocol ◽  
Factor Release

ABSTRACTAimA force of 400g at 2700 RPM results in an optimum leucocyte and platelet-rich fibrin (L-PRF). Most of centrifuges with varying characteristics generate a g-force in excess of 700g at 2700 RPM. In this context, the study explores the effect of the original centrifugation protocol and a modified protocol tailor-made to lower the RPM to generate a g-force of ~400g on platelet concentration, clot size and growth factors release in L-PRF prepared in two different commercially available centrifuges.Materials and Methods25 subjects each were assigned to the following groups; R1 and R2 where L-PRF was obtained from two laboratory swing-out centrifuges (Remi 8C® & Remi C854®, Mumbai, India) respectively. PRF was obtained from each subject within a group using two protocols; Original (O) protocol: conforming to the original centrifugation cycle (2700 RPM for 12 min) and Modified (M) protocol. Clot size, growth factor estimation and platelet counts were measured at 20, 40 and 60 mins from all the L-PRF clots.ResultsAt the third time period (40–60min), there were no significant differences in clot sizes with the original protocol (p=0.09), but a highly significant difference was noticed with the modified protocol in both the centrifuges (p=0.001). Our results showed an increased concentration of VEGF and EGF with modified protocol than with original protocol with both the centrifuges (p=0.001). By the end of second and third time periods, more platelet concentration was observed with modified protocol than with the original protocol in both the centrifuges (p=0.001)ConclusionThis study infers that the centrifuge type and RCF can affect the quality and quantity of cells and growth factors and an optimum relationship between g-force and RPM should be maintained in order to obtain L-PRF with adequate cell viability and optimum growth factor release.

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