Activated PRP versus PRGF (ie ENDORET®)

This is for a patient who asked what the difference is between PRP and PRGF.

The answer: it appears to be the same.
Platelet Rich Plasma as is typically used by most doctors and surgeon in the US.

The issue is the following:
1. PRP technically is just platelets and not activated with Calcium Chloride (see Ref 1 paper).
2. PRGF is activated PRP which means generally calcium choride was added as noted in below published protocol from Reference 1.

Activated PRP is what is used in the majority of PRP injection cases to my knowledge in the US.

Thus activated PRP is the same as PRGF.

It appears that the authors of the papers on PRGF or Endoret® had a significant vested interest. Either way activated PRP and PRGF do work to help patients with dry eye.
Be sure your surgeon is using activate PRP.

http://bti-biotechnologyinstitute.com/us/regenerative-medicine/applications-endoret/ophthalmology/

^{Reference 1:}

2012 Aug 15;53(9):5571-8. doi: 10.1167/iovs.11-7340.

In vitro effects of three blood derivatives on human corneal epithelial cells.

Freire V¹, Andollo N, Etxebarria J, Durán JA, Morales MC.

Author information

1: R&D&I Department, Instituto Clínico-Quirúrgico de Oftalmología, Bilbao, Spain.

Abstract

PURPOSE:

We compared the effects of three blood derivatives, autologous serum (AS), platelet-rich plasma (PRP), and serum derived from plasma rich in growth factors (PRGF), on a human corneal epithelial (HCE) cell line to evaluate their potential as an effective treatment for corneal epithelial disorders.

METHODS:

The concentrations of epidermal growth factor (EGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), and fibronectin were quantified by ELISA. The proliferation and viability of HCE cells were measured by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay. Cell morphology was assessed by phase-contrast microscopy. The patterns of expression of several connexin, involucrin, and integrin α6 genes were analyzed by real-time RT-PCR.

RESULTS:

We found significantly higher levels of EGF in PRGF compared to AS and PRP. However, AS and PRGF induced robust proliferation of HCE cells. In addition, PRGF cultured cells grew as heterogeneous colonies, exhibiting differentiated and non-differentiated cell phenotypes, whereas AS- and PRP-treated cultures exhibited quite homogeneous colonies. Finally, PRGF upregulated the expression of several genes associated with communication and cell differentiation, in comparison to AS or PRP.

CONCLUSIONS:

PRGF promotes biological processes required for corneal epithelialization, such as proliferation and differentiation. Since PRGF effects are similar to those associated with routinely used blood derivatives, the present findings warrant further research on PRGF as a novel alternative treatment for ocular surface diseases.

Blood from 16 healthy volunteers was collected by venipuncture (age range 30–60 years) after patient consent had been signed, in accordance with the Declaration of Helsinki. All volunteers were healthy and not taking any medication. The blood sample from each volunteer was processed according to the following three methods to obtain the corresponding blood derivatives:

Autologous serum (AS): Spontaneous coagulation for 2 hours at room temperature followed by centrifugation at 1,000 g for 15 minutes. Collection of the complete supernatant fraction.¹⁷
Platelet-rich plasma (PRP): Centrifugation at 460 g for 8 minutes and collection of the complete supernatant fraction.²⁴
Serum derived from plasma rich in growth factors (PRGF): Centrifugation at 460 g for 8 minutes, followed by collection of the complete supernatant fraction and induction of clot formation by adding calcium chloride at a final concentration of 22.8 mM (BTI Biotechnology Institute, S.L., Miñano, Álava, Spain), in the absence of red and white blood cells. After 2 hours at 36°C, the clot was retracted and the supernatant was collected.^28,29

Notes:

Serum derived from plasma rich in growth factors (PRGF): Centrifugation at 460 g for 8 minutes, followed by collection of the complete supernatant fraction and induction of clot formation by adding calcium chloride at a final concentration of 22.8 mM (BTI Biotechnology Institute, S.L., Miñano, Álava, Spain), in the absence of red and white blood cells. After 2 hours at 36°C, the clot was retracted and the supernatant was collected.^28,29

ttps://www.nature.com/articles/eye2017182
Correspondence

Searching for the best blood-derived eye drops

E Anitua^{1, 2, 3}
, R Prado ORCID: orcid.org/0000-0003-4767-7206^{1, 2, 3}
, F Muruzabal ORCID: orcid.org/0000-0001-7359-1698^{1, 2, 3}
[…]
& G Orive^{1, 2, 3}
– Show fewer authors

Sir,

We have read with great interest the article by Than et al¹ regarding the use of fingerprick autologous blood (FAB) as an alternative treatment for dry eye syndrome (DES). Sixteen patients diagnosed with DES were treated with FAB, 11 of whom had Sjögren syndrome, obtaining good results for the different clinical variables evaluated. The treatment dosage was four times daily for eight weeks.

We commend the authors’ search of a low-cost and readily accessible treatment for this type of syndrome. At the same time, we would like to offer some commentaries and additional perspectives to the study.¹

First, our experience is that, in order to achieve optimal therapeutic effects, it would be recommended that the concentrations of platelets and growth factors would be greater than those obtained in whole blood. A good example of that is platelet-rich plasma (PRP),² a therapy in which platelets are concentrated in a volume of plasma. Furthermore, we speculate that if platelet activation is not completed in FAB, the amount of platelet-derived growth factors would be lower than even autologous serum (AS).

Second, the authors should clarify whether there is platelet activation in the ocular surface, or whether it does not always occur, for example due to the clearance of FAB. The authors state that clots are observed in some patients, but this should be generalized so that the method is reproducible. Recent studies carried out on ocular surface cells show that the biological activity of non-activated PRP is lower than AS or other activated PRPs, such as plasma rich in growth factors (PRGF).³

Third, it is important to highlight that most of the patients (11/16) had been diagnosed with Sjögren syndrome, a long-term autoimmune disease in which the already exacerbated inflammatory component of DES is accentuated.⁴ An interesting approach for these cases might be to perform an inactivation protocol of immune components in the eye drops, preserving most of the biologically active molecules while reducing immunoglobulin content and complement activity.⁵

Last but not least, there are other issues to be considered, including the drawback of not using a standardized ready-to-use product, the large amount of fingerpricks delivered (448 fingerpricks in 8 weeks), and the variability of capillary blood.⁶

We strongly encourage the authors to further deepen research in the FAB mechanisms of action and product composition in order to optimize the treatment and improve the quality of life of DES patients, thus offering a fully characterized product.

ReferencesReferences

1.
Than J, Balal S, Wawrzynski J, Nesaratnam N, Saleh GM, Moore J et al. Fingerprick autologous blood: a novel treatment for dry eye syndrome. Eye (Lond) 2017; 31: 1655–1663.
2.
Anitua E, de la Fuente M, Muruzabal F, Riestra A, Merayo-Lloves J, Orive G. Plasma rich in growth factors (PRGF) eye drops stimulates scarless regeneration compared to autologous serum in the ocular surface stromal fibroblasts. Exp Eye Res 2015; 135: 118–126.
3.
Freire V, Andollo N, Etxebarria J, Hernaez-Moya R, Duran JA, Morales MC. Corneal wound healing promoted by 3 blood derivatives: an in vitro and in vivo comparative study. Cornea 2014; 33(6): 614–620.
4.
Barabino S, Chen Y, Chauhan S, Dana R. Ocular surface immunity: homeostatic mechanisms and their disruption in dry eye disease. Prog Retin Eye Res 2012; 31(3): 271–285.
- - CAS
  - PubMed
  - Article
  - Google Scholar
5.
Sanchez-Avila RM, Merayo-Lloves J, Riestra AC, Anitua E, Muruzabal F, Orive G et al. The effect of immunologically safe plasma rich in growth factor eye drops in patients with Sjogren syndrome. J Ocul Pharmacol Ther 2017; 33(5): 391–399.
6.
Bond MM, Richards-Kortum RR. Drop-to-drop variation in the cellular components of fingerprick blood: implications for point-of-care diagnostic development. Am J Clin Pathol 2015; 144(6): 885–894.

Download references

Author informationAuthor information

Affiliations

Foundation Eduardo Anitua, Vitoria, Spain
- E Anitua
- , R Prado
- , F Muruzabal
- & G Orive
Biotechnology Institute (BTI), Vitoria, Spain
- E Anitua
- , R Prado
- , F Muruzabal
- & G Orive
University Institute for Regenerative Medicine and Oral Implantology – UIRMI (UPV/EHU-Fundación Eduardo Anitua), Vitoria, Spain
- E Anitua
- , R Prado
- , F Muruzabal
- & G Orive

Competing interests

The authors declare that EA is the Scientific Director of and GO, RP, and FM are scientists at BTI Biotechnology Institute, a biomedical company that investigates in the fields of oral implantology and PRGF-Endoret technology.

2017 Jun;33(5):391-399. doi: 10.1089/jop.2016.0166. Epub 2017 Apr 4.

The Effect of Immunologically Safe Plasma Rich in Growth Factor Eye Drops in Patients with Sjögren Syndrome.

Sanchez-Avila RM¹, Merayo-Lloves J¹, Riestra AC¹, Anitua E^2,³, Muruzabal F^2,³, Orive G^3,^4,⁵, Fernández-Vega L¹.

Author information

1: 1 Instituto Universitario Fernández-Vega, Fundación de Investigación Oftalmológica, Universdiad de Oviedo, Oviedo, Spain .
2: 2 Fundación Eduardo Anitua, Vitoria, Spain .
3: 3 Biotechnology Institute (BTI) , Vitoria, Spain .
4: 4 NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country , School of Pharmacy, Vitoria, Spain .
5: 5 Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN), Vitoria, Spain .

Abstract

PURPOSE:

The objective was to provide preliminary information about the efficacy and safety of immunologically safe plasma rich in growth factor (immunosafe PRGF) eye drops in the treatment of moderate to severe dry eye in patients with primary and secondary Sjögren’s syndrome (SS) and to analyze the influence of several variables on treatment outcomes.

METHODS:

This retrospective study included patients with SS. All patients were treated with previously immunosafe PRGF eye drops to reduce the immunologic component contents. Ocular Surface Disease Index (OSDI) scale, best-corrected visual acuity (BCVA), visual analog scale (VAS) frequency, and VAS severity outcome measures were evaluated before and after treatment with immunosafe PRGF. The potential influence of some patient clinical variables on results was also assessed. Safety assessment was also performed reporting all adverse events.

RESULTS:

Twenty-six patients (12 patients with primary SS, and 14 patients suffering secondary SS) with a total of 52 affected eyes were included and evaluated. Immunosafe PRGF treatment showed a significant reduction (P < 0.05) in OSDI scale (41.86%), in BCVA (62.97%), in VAS frequency (34.75%), and in VAS severity (41.50%). BCVA and VAS frequency scores improved significantly (P < 0.05) after concomitant treatment of PRGF with corticosteroids. Only 2 adverse events were reported in 2 patients (7.7% of patients).

CONCLUSIONS:

Signs and symptoms of dry eye syndrome in patients with SS were reduced after treatment with PRGF-Endoret eye drops. Immunosafe PRGF-Endoret is safe and effective for treating patients with primary and secondary SS.

2015 Sep;34(9):1144-8. doi: 10.1097/ICO.0000000000000489.

Preservation of Biological Activity of Plasma and Platelet-Derived Eye Drops After Their Different Time and Temperature Conditions of Storage.

Anitua E¹, de la Fuente M, Riestra A, Merayo-Lloves J, Muruzábal F, Orive G.

Author information

Abstract

PURPOSE:

To analyze whether plasma rich in growth factors (PRGF) eye drops preserve their biological characteristics and activity after storage for 3 and 6 months at -20°C, at 4°C, and at room temperature for 72 hours, compared with fresh samples (t0).

METHODS:

Blood from 6 healthy donors was harvested and centrifuged to obtain PRGF free of leukocytes. Resulting PRGF eye drops were stored for 3 and 6 months at -20°C. At each time, 2 aliquots were maintained at room temperature or at 4°C for 72 hours. Platelet-derived growth factor-AB, transforming growth factor-β1, vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor-1, angiopoietin-1, and thrombospondin-1 were quantified at each time and temperature of storage. Also, the effect of PRGF eye drops on proliferation of primary human keratocytes was evaluated.

RESULTS:

All the analyzed growth factor levels remained constant at each time and storage condition. No differences were observed in the proliferative activity of keratocytes after treatment with PRGF eye drops at any studied time or temperature. Finally, there was no microbial contamination in any of the PRGF eye drops.

CONCLUSIONS:

The preservation of the PRGF eye drops at -20°C for up to 3 and 6 months does not mean reduction of the main growth factors and proteins implicated in ocular surface wound healing. Eye drop characteristics and in vitro biological activity were not affected by their usage and conservation for 72 hours at 4°C or at room temperature.

PRGF Sample Preparation PRGF eye drops from each donor were obtained using the Endoret-Ophthalmology kit (BTI Biotechnology Institute, SL, Miñano, Álava, Spain).

Plasma rich in growth factors (PRGF) is an autologous platelet-rich plasma based on the use of the patient’s own blood, and the resulting therapeutic formulations derived from this technology contain a wide variety of growth factors, proteins, and biomaterials involved in tissue regeneration. The effectiveness, signiﬁcant clinical advantages, and safety of this approach have been deeply demonstrated in several medical ﬁelds.2–5 In fact, PRGF provides a pool of biologically active proteins and growth factors, including plateletderived growth factor-AB (PDGF-AB), transforming growth factor-b1 (TGF-b1), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), thrombospondin-1 (TSP-1), and angiopoietin-1 (ANG-1) among others. These proteins are involved in a range of biological processes, including cell recruitment, growth, and differentiation.6,7

The authors declare the following competing financial interest(s): E.A. is the Scientific Director of and G.O. and F.M. are scientists at BTI Biotechnology Institute, a dental implant company that investigates in the fields of oral implantology and PRGF-Endoret technology

http://bti-biotechnologyinstitute.com/us/regenerative-medicine/applications-endoret/ophthalmology/

In vitro effects of three blood derivatives on human corneal epithelial cells.

Author information

Abstract

PURPOSE:

METHODS:

RESULTS:

CONCLUSIONS:

Searching for the best blood-derived eye drops

ReferencesReferences

Author informationAuthor information

Affiliations

Foundation Eduardo Anitua, Vitoria, Spain

Biotechnology Institute (BTI), Vitoria, Spain

University Institute for Regenerative Medicine and Oral Implantology – UIRMI (UPV/EHU-Fundación Eduardo Anitua), Vitoria, Spain

Competing interests

The Effect of Immunologically Safe Plasma Rich in Growth Factor Eye Drops in Patients with Sjögren Syndrome.

Author information

Abstract

PURPOSE:

METHODS:

RESULTS:

CONCLUSIONS:

Preservation of Biological Activity of Plasma and Platelet-Derived Eye Drops After Their Different Time and Temperature Conditions of Storage.

Author information

Abstract

PURPOSE:

METHODS:

RESULTS:

CONCLUSIONS: