scholarly journals Comparison of Two Preparation Methods for Platelet-Rich Plasma Eye Drops for Release of Growth Factors and De-Epithelization Rabbit Model

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Tatsuhiko Kobayashi ◽  
Takashi Suzuki ◽  
Tomohiko Saito ◽  
Takashi Itokawa ◽  
Yuichi Hori

Purpose. To compare two platelet-rich plasma (PRP) preparation methods (double spin (D-PRP) and TriCell PRP (T-PRP)) for eye drops, concentration yields of platelets and other cells, release of growth factors, and efficacy of the de-epithelization rabbit model. Methods. PRP was extracted by D-PRP and T-PRP from 30 ml blood from healthy adults. After extraction, platelets and leukocytes were counted. D-PRP and T-PRP were preserved during A: 1 h storage at room temperature, B: 1 h storage at −20°C, C: 24 h storage at 4°C, and D: 24 h storage at −20°C. Platelet-derived growth factor (PDGF) was measured. Freezing/thawing PRP eye drops and control were instilled in the de-epithelization rabbit model four times per day for 5 days. Histology was compared between eyes treated with control, D-PRP, and T-PRP. Results. 14 ml of D-PRP and 4 ml of T-PRP were extracted from 30 ml whole blood samples. D-PRP and T-PRP had 41.36 ± 8.43 × 104 and 67.02 ± 13.55 × 104 platelets and 0.41 ± 0.24 × 103/ml and 10.09 ± 4.29 × 103/ml leucocytes, respectively. In the four storage methods, PDGF concentrations in T-PRP were higher than those in D-PRP eye drops. Freezing/thawing D-PRP and T-PRP increased PDGF concentrations. Histology showed corneal epithelium thickness in T-PRP-treated eyes after healing (38.41 ± 9.10 μm) was significantly higher than that in control-treated (27.77 ± 4.76 μm) and D-PRP-treated eyes (18.32 ± 3.14 μm) ( P  < 0.05). There was no corneal damage with inflammation in corneal stroma and epithelium of all tested groups. Electron microscopy revealed strong adhesion between cell junctions in T-PRP-treated eyes. Conclusions. Freezing/thawing of PRP extracted with the T-PRP kit may result in high platelet and leukocyte concentration and produce high PDGF concentration. PRP eye drops including leucocytes could increase thickness of corneal epithelium without corneal inflammation.

2018 ◽  
Vol 59 (3-4) ◽  
pp. 265-275 ◽  
Author(s):  
Siegmund Lang ◽  
Markus Loibl ◽  
Marietta Herrmann

Background: Platelet-rich plasma (PRP) refers to an enriched platelet suspension in plasma. In addition to the clinical application of PRP in the context of various orthopedic diseases and beyond, PRP and platelet lysate (PL) have been in focus in the field of tissue engineering. In this review, we discuss the application of PRP as a cell culture supplement and as part of tissue engineering strategies, particularly emphasizing current hurdles and ambiguities regarding the efficacy of PRP in these approaches. Summary: As a putative autologous replacement for animal-derived supplements such as fetal calf serum (FCS), PRP has been applied as cell culture supplement for the expansion of stem and progenitor cells for tissue engineering applications and cell therapies. Attributed to the high content of growth factors in platelets, PRP has been shown to promote cell growth, which was mostly superior to standard cultures supplemented with FCS, while the differentiation capacity of progenitor cells seems not to be affected. However, it was also suggested that cultivation of cells with PRP significantly alters the protein expression profile in cells in comparison to FCS, indicating that the influence of PRP on cell behavior should be thoroughly investigated. Moreover, different PRP preparation methods and donor variations have to be considered for the use of PRP under good manufacturing practice conditions. PRP has been used for various tissue engineering applications in the context of bone, cartilage, skin, and soft tissue repair, where most studies were conducted in the field of bone tissue engineering. These approaches take either advantage of the release of chemoattractive, angiogenic, proliferative, and putatively pro-regenerative growth factors from PRP, and/or the hydrogel properties of activated PRP, making it suitable as a cell delivery vehicle. In many of these studies, PRP is combined with biomaterials, cells, and in some cases recombinant growth factors. Although the experimental design often does not allow conclusions on the pro-regenerative effect of PRP itself, most publications report beneficial effects if PRP is added to the tissue-engineered construct. Furthermore, it was demonstrated that the release of growth factors from PRP may be tailored and controlled when PRP is combined with materials able to capture growth factors. Key Messages: Platelet-derived preparations such as PRP and PL represent a promising source of autologous growth factors, which may be applied as cell culture supplement or to promote regeneration in tissue-engineered constructs. Furthermore, activated PRP is a promising candidate as an autologous cell carrier. However, the studies investigating PRP in these contexts often show conflicting results, which most likely can be attributed to the lack of standardized preparation methods, particularly with regard to the platelet content and donor variation of PRP. Ultimately, the use of PRP has to be tailored for the individual application.


2016 ◽  
Vol 61 (No. 10) ◽  
pp. 567-570 ◽  
Author(s):  
J. Vokurka ◽  
E. Gopfert ◽  
M. Blahutkova ◽  
E. Buchalova ◽  
M. Faldyna

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Johannes Zellner ◽  
Christian Dirk Taeger ◽  
Markus Schaffer ◽  
J. Camilo Roldan ◽  
Markus Loibl ◽  
...  

Meniscal lesions in the avascular zone are still a problem in traumatology. Tissue Engineering approaches with mesenchymal stem cells (MSCs) showed successful regeneration of meniscal defects in the avascular zone. However, in daily clinical practice, a single stage regenerative treatment would be preferable for meniscus injuries. In particular, clinically applicable bioactive substances or isolated growth factors like platelet-rich plasma (PRP) or bone morphogenic protein 7 (BMP7) are in the focus of interest. In this study, the effects of PRP and BMP7 on the regeneration of avascular meniscal defects were evaluated. In vitro analysis showed that PRP secretes multiple growth factors over a period of 8 days. BMP7 enhances the collagen II deposition in an aggregate culture model of MSCs. However applied to meniscal defects PRP or BMP7 in combination with a hyaluronan collagen composite matrix failed to significantly improve meniscus healing in the avascular zone in a rabbit model after 3 months. Further information of the repair mechanism at the defect site is needed to develop special release systems or carriers for the appropriate application of growth factors to support biological augmentation of meniscus regeneration.


2015 ◽  
Vol 28 (03) ◽  
pp. 164-171 ◽  
Author(s):  
J. Schneppendahl ◽  
T. T. Lögters ◽  
M. Sager ◽  
M. Wild ◽  
M. Hakimi ◽  
...  

Summary Introduction: Large bone defects are a therapeutic challenge to surgeons and are often associated with a high morbidity. The use of autologous cancellous bone graft represents an essential therapeutic option and is considered the gold standard. However, the use of platelet-rich plasma (PRP) for improving bone defect healing has been discussed controversially. The aim of this study was to evaluate the treatment of a diaphyseal long-bone defect in a rabbit model with a combination of PRP and autologous cancellous bone. Material and methods: A monocortical long-bone defect in the radial diaphysis of 24 New Zealand white rabbits was filled either with autologous cancellous graft as a control group or with autologous cancellous graft combined with autologous PRP. Histomorphometrical and radiological analysis as well as quantification of platelets and growth factors were performed. The animals were euthanatized after three and six weeks according to the study arms. Results: A significant improvement in bone healing was observed histomorphometrically in the PRP group in the central area of the defect zone (p <0.01) as well as the cortical defect zone (p <0.01). The radiological findings were in accordance with the histomorphometrical results. Comparing native blood and PRP, an enrichment of growth factors and platelets was detectable in the PRP. Conclusion: Within this animal study, the combination of PRP and autologous cancellous bone grafts improved bone healing significantly compared to the sole application of autologous bone. Therefore, further efforts should be initiated to establish the composite of PRP and autologous bone for bone healing disorders in clinical use.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Chee Wai Wong ◽  
Ning Cheung ◽  
Candice Ho ◽  
Veluchamy Barathi ◽  
Gert Storm ◽  
...  

Abstract To clarify the mechanisms and their temporal relationship in the development of proliferative vitreoretinopathy (PVR), we measured vitreous levels of pro-inflammatory cytokines and growth factors in a rabbit model of PVR. PVR was surgically induced in 11 rabbit eyes by vitrectomy, retinotomy, cryotherapy and injection of platelet-rich plasma at baseline. Severity of PVR was assessed on dilated fundal examination with indirect binocular ophthalmoscopy and graded based on the revised experimental PVR classification. Severe PVR was defined as stage 5 or worse. Vitreous concentrations of interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 1 beta (IL-1 β), tumor necrosis factor beta (TNF-β), granulocyte macrophage colony stimulating factor (GM-CSF), interferon gamma (IFN-γ), C reactive protein; (CRP), placental growth factor (PlGF), platelet derived growth factor BB (PDGF-BB), vascular endothelial growth factor (VEGF) and angiopoietin 2 (Ang-2) at weeks 2, 3 and 4 were compared to baseline and correlations between the cytokines with PVR severity were assessed. Four weeks after PVR induction, 5 eyes (45.5%) had developed severe PVR. IL-8 was raised at 2 weeks post PVR induction (1.46 ± 0.48 pg/ml vs 0.53 ± 0.25 pg/ml, p = 0.04) and remained significantly elevated at week 4 (2.6 ± 3.1 pg/ml, p = 0.03). CRP was significantly raised at week 4 (34.8 ± 12.0 pg/ml vs 13.0 ± 13.1 pg/ml, p < 0.001). Among the growth factors, PDGF-BB was the earliest to show significantly elevated levels, at 3 weeks (50.4 ± 19.0 pg/ml vs 6.2 ± 10.1 pg/ml) and remained elevated at week 4 (p = 0.002), while PlGF (11.2 ± 7.7 pg/ml vs 5.3 ± 3.8 pg/ml, p = 0.002) and Ang2 (13617.0 ± 8170.2 pg/ml vs 38593.8 ± 8313.4, p = 0.02) were significantly raised at week 4. IFN-γ (p = 0.03), PDGF-BB (p = 0.02) and VEGF (p = 0.02) were significantly associated with PVR severity. We demonstrated that inflammatory cytokines IL-6, -8, elevation post PVR induction is followed by elevated levels of fibroproliferative growth factors, Ang2, PlGF, VEGF and PDGF-BB in the development of PVR. These findings will guide future studies targeting appropriate therapeutic strategies for the treatment of PVR.


2020 ◽  
Vol 23 (4) ◽  
pp. 252-257
Author(s):  
A. A. Lepehova ◽  
Maria B. Potapova

Androgenetic alopecia is a genetically determined disease characterized by gradual thinning of hair, which subsequently leads to hair loss, and thereby significantly affects a patients quality of life. The existing methods of disease management have a number of side effects and drawbacks. Thus, new treatment methods combining both high tolerance and efficacy are continuously determined. This current review presents an analysis of the effectiveness of single- and double-spin preparation methods of platelet-rich plasma for the management of patients with androgenetic alopecia.


Biology ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 317
Author(s):  
Eitan Mijiritsky ◽  
Haya Drora Assaf ◽  
Oren Peleg ◽  
Maayan Shacham ◽  
Loredana Cerroni ◽  
...  

Growth factors (GFs) play a vital role in cell proliferation, migration, differentiation and angiogenesis. Autologous platelet concentrates (APCs) which contain high levels of GFs make them especially suitable for periodontal regeneration and facial rejuvenation. The main generations of APCs presented are platelet-rich plasma (PRP), platelet-rich fibrin (PRF) and concentrated growth factor (CGF) techniques. The purpose of this review is to provide the clinician with an overview of APCs’ evolution over the past decade in order to give reliable and useful information to be used in clinical work. This review summarizes the most interesting and novel articles published between 1997 and 2020. Electronic and manual searches were conducted in the following databases: Pubmed, Scopus, Cochrane Library and Embase. The following keywords were used: growth factors, VEGF, TGF-b1, PRP, PRF, CGF and periodontal regeneration and/or facial rejuvenation. A total of 73 articles were finally included. The review then addresses the uses of the three different techniques in the two disciplines, as well as the advantages and limitations of each technique. Overall, PRP is mainly used in cases of hard and soft tissue procedures, while PRF is used in gingival recession and the treatment of furcation and intrabony defects; CGF is mainly used in bone regeneration.


Vision ◽  
2021 ◽  
Vol 5 (3) ◽  
pp. 34
Author(s):  
Ronald M. Sánchez-Ávila ◽  
Edmar Uribe-Badillo ◽  
Carlos Fernández-Vega González ◽  
Francisco Muruzabal ◽  
Borja de la Sen-Corcuera ◽  
...  

This study aimed to investigate the use of Plasma Rich in Growth Factors (PRGF) associated with tissue ReGeneraTing Agent (RGTA) drops for the treatment of noninfectious corneal ulcers. RGTA treatment was applied (one drop every two days); however, if ulcer closure was not achieved, PRGF eye drops treatment was added (four times/day). The time taken to reach the ulcer closure, the Best Corrected Visual Acuity (BCVA), intraocular pressure (IOP), Visual Analog Scale (VAS, in terms of frequency and severity of symptoms), and Ocular Surface Disease Index (OSDI) were evaluated. Seventy-four patients (79 eyes) were included, and the mean age was 56.8 ± 17.3 years. The neurotrophic corneal ulcer was the most frequent disorder (n = 27, 34.2%), mainly for herpes virus (n = 15, 19.0%). The time of PRGF eye drops treatment associated with the RGTA matrix was 4.2 ± 2.2 (1.5–9.0) months, and the follow-up period was 44.9 ± 31.5 months. The ulcer closure was achieved in 76 eyes (96.2%). BCVA, VAS and OSDI improved from the baseline (p < 0.001), and IOP remained unchanged (p = 0.665). RGTA and PRGF in noninfectious ulcers were effective and could be a therapeutic alternative for this type of corneal disease.


2021 ◽  
Vol 14 (8) ◽  
pp. 753
Author(s):  
Anna Matysik-Woźniak ◽  
Waldemar A. Turski ◽  
Monika Turska ◽  
Roman Paduch ◽  
Mirosław Łańcut ◽  
...  

Kynurenic acid (KYNA) is an endogenous compound with a multidirectional effect. It possesses antiapoptotic, anti-inflammatory, and antioxidative properties that may be beneficial in the treatment of corneal injuries. Moreover, KYNA has been used successfully to improve the healing outcome of skin wounds. The aim of the present study is to evaluate the effects of KYNA on corneal and conjunctival cells in vitro and the re-epithelization of corneal erosion in rabbits in vivo. Normal human corneal epithelial cell (10.014 pRSV-T) and conjunctival epithelial cell (HC0597) lines were used. Cellular metabolism, cell viability, transwell migration, and the secretion of IL-1β, IL-6, and IL-10 were determined. In rabbits, after corneal de-epithelization, eye drops containing 0.002% and 1% KYNA were applied five times a day until full recovery. KYNA decreased metabolism but did not affect the proliferation of the corneal epithelium. It decreased both the metabolism and proliferation of conjunctival epithelium. KYNA enhanced the migration of corneal but not conjunctival epithelial cells. KYNA reduced the secretion of IL-1β and IL-6 from the corneal epithelium, leaving IL-10 secretion unaffected. The release of all studied cytokines from the conjunctival epithelium exposed to KYNA was unchanged. KYNA at higher concentration accelerated the healing of the corneal epithelium. These favorable properties of KYNA suggest that KYNA containing topical pharmaceutical products can be used in the treatment of ocular surface diseases.


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