Freeze-dried versus fresh platelet-rich plasma in acute wound healing of an animal model

Abstract Background: Increasing prevalence of diabetes and obesity has registered a simultaneous increase in chronic wounds and defects in tissue repair. Recently PRP was described to be used in the treatment of chronic ulcers. PRP contains cytokines, chemokines and growth factors (GF) derived from platelets which induce molecular and cell mechanisms of natural wound healing. The possible use of GF derived from platelets, and the potential to prolong the shelf life of platelet concentrates, freeze-drying becomes one particularly suitable method to improve wound healing treatment. The use of lyophilized PRP appears to be interesting for greater convenience: only one blood collection, after hydration its soon ready for use, and also due to the highest concentration of bioactive molecules. This study aims to compare intra-lesional freeze-dried and fresh PRP for treatment of acute wound in a pre-clinic model. Methods: Fresh and freeze-dried PRP was prepared from two human platelet concentrate bags using double spin method. PRP characterization included platelet number and quantification of PDGF-AA, EGF, VEGF and TGF-β1 by Luminex (Millipore). Animal wound model was performed after shaving the dorsum of the rat, and a full-thickness excisional wound (1 cm2) was made to the level of the panniculus carnosus muscle. Thirty animals were divided according to the type o treatment in 3 groups: Fresh PRP (n=10), Freeze-dried PRP (FD-PRP) (n=10) and control saline (n=10). The animals received one perilesional application of 500 µL in the day of wound induction. The monitoring of wound closure was made through macroscopical analysis of wound size in days 3, 7 and 10 after wounding. Animals were sacrificed in the tenth day and the skin was removed for histology with hematoxilin-eosin and Masson´s Trichrome, and immunohistochemistry with α-actin smooth muscle for myofibroblasts and blood vessels, and quantified through image J. Results: The platelet number of PRP was 5714 x 103 cells / µL, presenting a high platelet recovery. The FD-PRP presented higher level of all GF when compared to fresh PRP from 1.64 to 3.72 folds. The application of PRP or FD-PRP did not induce significantly changes in wound healing kinetics compared to control during all evaluated days. The mean and standard deviation (SD) of area in D3 was 112.9 ± 16.6%, 107.9 ± 21.3 and 100.8 ± 30.2 for control, PRP and FD-PRP, respectively. In D7 it was 54.3 ± 28.9, 63.9 ± 22.8 and 61.6 ± 20.6 for control, PRP and FD-PRP, respectively. In D10 it was 9.9 ± 6.1, 13.9 ± 11.2, 7.7 ± 6.0 for control, PRP and FD-PRP, respectively. In immunohistochemistry of deep epidermis FD-PRP presented a significantly higher concentration of myofibroblasts in comparison with fresh PRP (16.61 ± 9.04 vs. 13.99 ± 14.07, p=0.0095). A significantly higher number of blood vessels was observed in the group treated with FD-PRP (in percentage of area) in comparison to the controls, both in superficial and deep regions of epidermis (0.43 ± 0.5 vs. 0.21 ± 0.22, P=0.01; 0.38 ± 0.44 vs. 0.24 ± 0.25, P=0.03, respectively). Discussion and conclusion: The most interesting result of this study was the increased number of blood vessels. VEGF plays a central role in promoting angiogenesis during wound repair. Previous studies demonstrated accelerated wound closure with collagen-binding VEGF topical treatment. However, the use of one growth factor has limited success and heterogeneity of clinical results, probably due to the need of multiple bioactive molecules that are necessary in the cascade of complexes events of healing. The use of multiples factors necessary to healing can orchestrated the physiological events resulting in chemotaxis, proliferation and differentiation of cells and the angiogenic process. The use of FD-PRP in this study of acute model of wounds showed positive results, especially in angiogenesis. The practicality and the improve in the shelf life of PRP have a great value in clinical practice and becomes very attractive, showing low cost, the possibility to increase the concentration of GF, various samples ready-to-use and a single blood collection. The investigation of FD-PRP in other clinical situations in which fresh PRP have been used is an interesting approach to be to evaluated to determine its effectiveness. Figure Quantification of blood vessels through imunohistochemistry using smooth α-actin and treated by Image J software; A) Superficial epidermis; B) Deep epidermis. Figure. Quantification of blood vessels through imunohistochemistry using smooth α-actin and treated by Image J software; A) Superficial epidermis; B) Deep epidermis. Disclosures No relevant conflicts of interest to declare.

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Changes in optical properties of tissue during acute wound healing in an animal model

Journal of Biomedical Optics ◽

10.1117/1.2960952 ◽

2008 ◽

Vol 13 (4) ◽

pp. 044005 ◽

Cited By ~ 14

Author(s):

Elisabeth S. Papazoglou ◽

Michael S. Weingarten ◽

Leonid Zubkov ◽

Michael Neidrauer ◽

Linda Zhu ◽

...

Keyword(s):

Wound Healing ◽

Animal Model ◽

Optical Properties ◽

Acute Wound ◽

Optical Properties Of Tissue

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Allogenous Freeze-Dried Platelet-Rich Plasma Promotes Peripheral Myelin Repair in Chronic Constriction Nerve Injury

Systematic Reviews in Pharmacy ◽

10.31838/srp.2020.6.40 ◽

2020 ◽

Vol 11 (06) ◽

Keyword(s):

Nerve Injury ◽

Platelet Rich Plasma ◽

Myelin Repair ◽

Freeze Dried

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The evaluation of bone union activity of freeze-dried platelet-rich plasma

Bone Reports ◽

10.1016/j.bonr.2021.100909 ◽

2021 ◽

Vol 14 ◽

pp. 100909

Author(s):

Hideyuki Kinoshita ◽

Sumihisa Orita ◽

Yasuhiro Shiga ◽

Kazuhide Inage ◽

Seiji Ohtori

Keyword(s):

Platelet Rich Plasma ◽

Bone Union ◽

Union Activity ◽

Freeze Dried

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Mandibular Third Molar Extraction Wound Healing With and Without Platelet Rich Plasma: A Comparative Prospective Study

Journal of Maxillofacial and Oral Surgery ◽

10.1007/s12663-014-0738-1 ◽

2015 ◽

Vol 14 (3) ◽

pp. 808-815 ◽

Cited By ~ 9

Author(s):

Shubha Ranjan Dutta ◽

Purnima Singh ◽

Deepak Passi ◽

Pradeep Patter

Keyword(s):

Wound Healing ◽

Prospective Study ◽

Platelet Rich Plasma ◽

Third Molar ◽

Third Molar Extraction ◽

Mandibular Third Molar ◽

Comparative Prospective Study ◽

Molar Extraction

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Systematic Review: Adipose-Derived Mesenchymal Stem Cells, Platelet-Rich Plasma and Biomaterials as New Regenerative Strategies in Chronic Skin Wounds and Soft Tissue Defects

International Journal of Molecular Sciences ◽

10.3390/ijms22041538 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1538 ◽

Cited By ~ 2

Author(s):

Pietro Gentile ◽

Simone Garcovich

Keyword(s):

Systematic Review ◽

Stem Cells ◽

Wound Healing ◽

Soft Tissue ◽

Chronic Wounds ◽

Platelet Rich Plasma ◽

Skin Wounds ◽

Soft Tissue Defects ◽

Chronic Skin ◽

Tissue Defects

The number of clinical trials evaluating adipose-derived mesenchymal stem cells (AD-MSCs), platelet-rich plasma (PRP), and biomaterials efficacy in regenerative plastic surgery has exponentially increased during the last ten years. AD-MSCs are easily accessible from various fat depots and show intrinsic plasticity in giving rise to cell types involved in wound healing and angiogenesis. AD-MSCs have been used in the treatment of soft tissue defects and chronic wounds, employed in conjunction with a fat grafting technique or with dermal substitute scaffolds and platelet-rich plasma. In this systematic review, an overview of the current knowledge on this topic has been provided, based on existing studies and the authors’ experience. A multistep search of the PubMed, MEDLINE, Embase, PreMEDLINE, Ebase, CINAHL, PsycINFO, Clinicaltrials.gov, Scopus database, and Cochrane databases has been performed to identify papers on AD-MSCs, PRP, and biomaterials used in soft tissue defects and chronic wounds. Of the 2136 articles initially identified, 422 articles focusing on regenerative strategies in wound healing were selected and, consequently, only 278 articles apparently related to AD-MSC, PRP, and biomaterials were initially assessed for eligibility. Of these, 85 articles were excluded as pre-clinical, experimental, and in vitro studies. For the above-mentioned reasons, 193 articles were selected; of this amount, 121 letters, expert opinions, commentary, and editorials were removed. The remaining 72 articles, strictly regarding the use of AD-MSCs, PRP, and biomaterials in chronic skin wounds and soft tissue defects, were analyzed. The studies included had to match predetermined criteria according to the patients, intervention, comparator, outcomes, and study design (PICOS) approach. The information analyzed highlights the safety and efficacy of AD-MSCs, PRP, and biomaterials on soft tissue defects and chronic wounds, without major side effects.

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In vitro and in vivo Effect of Platelet-Rich Plasma-Based Autologous Topical Serum on Cutaneous Wound Healing

Skin Pharmacology and Physiology ◽

10.1159/000517195 ◽

2021 ◽

pp. 1-13

Author(s):

Eduardo Anitua ◽

Victoria Muñoz ◽

Libe Aspe ◽

Roberto Tierno ◽

Adrian García-Salvador ◽

...

Keyword(s):

Wound Healing ◽

Tissue Damage ◽

Collagen Synthesis ◽

Dermal Fibroblast ◽

Platelet Rich Plasma ◽

Cell Activity ◽

Cutaneous Wound Healing ◽

Cutaneous Wound ◽

Fibroblast Migration ◽

Edge Contraction

Introduction: Skin injury and wound healing is an inevitable event during lifetime. However, several complications may hamper the regeneration of the cutaneous tissue and lead to a chronic profile that prolongs patient recovery. Platelet-rich plasma is rising as an effective and safe alternative to the management of wounds. However, this technology presents some limitations such as the need for repeated blood extractions and health-care interventions. Objective: The aim of this study was to assess the use of an endogenous and storable topical serum (ES) derived from plasma rich in growth factors promoting wound healing, and to obtain preliminary data regarding its clinical and experimental effect over ulcerated skin models and patient care. Methods: Human dermal fibroblast and 3D organotypic ulcerated skin models were used to assess ES over the main mechanisms of wound healing including cell migration, edge contraction, collagen synthesis, tissue damage, extracellular matrix remodeling, cell death, metabolic activity, and histomorphometry analysis. Additionally, 4 patients suffering from skin wounds were treated and clinically assessed. Results: ES promoted dermal fibroblast migration, wound edge contraction, and collagen synthesis. When topically applied, ES increased collagen and elastin deposition and reduced tissue damage. The interstitial edema, structural integrity, and cell activity were also maintained, and apoptotic levels were reduced. Patients suffering from hard-to-heal wounds of different etiologies were treated with ES, and the ulcers healed completely within few weeks with no reported adverse events. Conclusion: This preliminary study suggests that ES might promote cutaneous wound healing and may be useful for accelerating the re-epithelization of skin ulcers.

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