Apheresis Platelet Rich-Plasma for Regenerative Medicine: An In Vitro Study on Osteogenic Potential

Background: Platelet-Rich Plasma (PRP) induces bone regeneration; however, there is low evidence supporting its efficacy in bone healing. The lack of a standardized protocol of administration represents the main obstacle to its use in the clinical routine for bone defects’ treatment. The purpose of this study was to characterize PRP and elucidate its osteogenic potential. Methods: Platelet count, fibrinogen levels, and growth factors concentration were measured in PRP obtained by four apheresis procedures. HOB-01-C1, a pre-osteocytic cell line, was used to examine the effects of different PRP dilutions (from 1% to 50%) on cell viability, growth, and differentiation. Gene expression of RUNX2, PHEX, COL1A1, and OCN was also assayed. Results: PRP showed a mean 4.6-fold increase of platelets amount compared to whole blood. Among the 36 proteins evaluated, we found the highest concentrations for PDGF isoforms, EGF, TGF-β and VEGF-D. PDGF-AA positively correlated with platelet counts. In three of the four tested units, 25% PRP induced a growth rate comparable to the positive control (10% FBS); whereas, for all the tested units, 10% PRP treatment sustained differentiation. Conclusions: This study showed that PRP from apheresis stimulates proliferation and differentiation of pre-osteocyte cells through the release of growth factors from platelets.

Download Full-text

In vitro study of the role of thrombin in platelet rich plasma (PRP) preparation: utility for gel formation and impact in growth factors release

Journal of Stem Cells and Regenerative Medicine ◽

10.46582/jsrm.1201002 ◽

2016 ◽

Vol 12 (1) ◽

pp. 2-9 ◽

Cited By ~ 12

Keyword(s):

Growth Factors ◽

Platelet Rich Plasma ◽

In Vitro Study ◽

Vitro Study ◽

Gel Formation

Download Full-text

In Vitro Studies on the Degradability, Bioactivity, and Cell Differentiation of PRP/AZ31B Mg Alloys Composite Scaffold

BioMed Research International ◽

10.1155/2017/5763173 ◽

2017 ◽

Vol 2017 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Jian Zou ◽

Zhongmin Shi ◽

Hongwei Xu ◽

Xiaolin Li

Keyword(s):

Water Absorption ◽

Composite Scaffold ◽

Platelet Rich Plasma ◽

In Vitro Study ◽

Mg Alloys ◽

Bone Tissue Regeneration ◽

Cell Seeding ◽

Proliferation And Differentiation ◽

Better Than

In recent years, more and more methods have been developed to improve the bioactivity of the biodegradable materials in bone tissue regeneration. In present study, we used rat mesenchymal stem cells (rMSCs) to evaluate the outcomes of Mg alloys (AZ31B, Magnesium, and Aluminum) and Platelet-rich plasma (PRP)/Mg alloys on rMSCs biocompatibility and osteogenic differentiation. Water absorption experiments indicated that both bare AZ31B and PRP/AZ31B were capable of absorbing large amounts of water. But the water absorption ratio for PRP/AZ31B was significantly higher than that for bare AZ31B. The degradability experiments implied that both samples degraded at same speed. rMSCs on the surface of AZ31B distributed more and better than those on the AZ31B scaffold. In ALP activity experiment, the activity of rMSCs on the PRP/AZ31B was markedly higher than that on the AZ31B scaffolds on the 7th day and 14th day. qRT-PCR also showed that OPN and OCN were expressed in both samples. OPN and OCN expression in PRP/AZ31B sample were higher than those in bare AZ31B samples. In summary, the in vitro study implied that AZ31B combined with PRP could remarkably improve cell seeding, attachment, proliferation, and differentiation.

Download Full-text

Effect of platelet-rich plasma concentrations on the proliferation of periodontal cells: An in vitro study

European Journal of Dentistry ◽

10.4103/1305-7456.195165 ◽

2016 ◽

Vol 10 (04) ◽

pp. 469-474 ◽

Cited By ~ 7

Author(s):

Sara Tavassoli-Hojjati ◽

Mandana Sattari ◽

Tayebeh Ghasemi ◽

Rahil Ahmadi ◽

Abbas Mashayekhi

Keyword(s):

Platelet Rich Plasma ◽

Plasma Concentrations ◽

In Vitro Study ◽

Positive Control ◽

Negative Control ◽

Repeated Measure ◽

Control Groups ◽

Healthy Human ◽

Significant Difference

ABSTRACT Objective: The purpose of this study was to evaluate the effect of different concentrations of platelet-rich plasma (PRP) on the proliferation of undifferentiated periodontal ligament (PDL) fibroblasts. Materials and Methods: The undifferentiated PDL fibroblasts were obtained from two healthy human premolar teeth and cultured in Dulbecco's modified Eagle's medium. Cell wells were divided into five groups. Experimental groups received 0.1%, 5%, or 50% PRP; the positive and negative control groups were cultured in fetal bovine serum (FBS) 12% and in a medium without FBS 12%, respectively. The plates were incubated at 37°C for 1, 2, 3, 4, and 7 days. PDL cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide proliferation assay. Statistical analysis of the data was accomplished using repeated measure ANOVA and Tukey's test. P < 0.05 was considered statistically significant. Results: The 5% PRP had the greatest effect on undifferentiated fibroblast proliferation, which was significant on the 3rd day. There was no significant difference between 0.1% PRP and positive control during the first 3 days. The group with 50% PRP presented significantly lower proliferation, compared to other experimental and control groups. Conclusions: It may be concluded that the growth-stimulating effect of PRP is dose dependent with the best results in low concentrations.

Download Full-text

Platelet-Rich Plasma Promotes Migration, Proliferation, and the Gene Expression of Scleraxis and Vascular Endothelial Growth Factor in Paratenon-Derived Cells In Vitro

Sports Health A Multidisciplinary Approach ◽

10.1177/1941738118807479 ◽

2018 ◽

Vol 11 (2) ◽

pp. 142-148 ◽

Cited By ~ 2

Author(s):

Sosuke Imai ◽

Ken Kumagai ◽

Yasuteru Yamaguchi ◽

Kazuma Miyatake ◽

Tomoyuki Saito

Keyword(s):

Gene Expression ◽

Platelet Rich Plasma ◽

Tendon Repair ◽

Tendon Healing ◽

Fold Increase ◽

Control Group ◽

Proliferation And Differentiation ◽

Fetal Bovine ◽

Endothelial Growth Factor

Background: Platelet-rich plasma (PRP) is a treatment option for tendon injury because of its effective tendon-healing properties. At the early stage of tendon repair, paratenon-derived cells (PDCs) are thought to play a more important role than tendon proper–derived cells (TDCs). However, there has been no study investigating the effects of PRP on PDCs. Hypothesis: PRP promotes the migration, proliferation, and differentiation of PDCs in vitro. Study Design: Controlled laboratory study. Methods: TDCs and PDCs were isolated from the tendon proper and paratenon of rat Achilles tendons and were cultured to the third passage. PRP was prepared from the rats using the double-spin method. Third-passage TDCs and PDCs were cultured in Dulbecco’s modified Eagle medium with 2% fetal bovine serum (control group) or 2% fetal bovine serum plus 5% PRP (PRP group), and cell migration, proliferation, and differentiation were evaluated. The relative mRNA expression levels of scleraxis (Scx), tenomodulin (Tnmd), collagen type I alpha 1 (Col1a1), collagen type III alpha 1 (Col3a1), and vascular endothelial growth factor A (VEGF) were examined by quantitative real-time reverse transcription polymerase chain reaction. Results: The cell migration rate was significantly higher in the PDCs of the PRP group than in the control group (1.4-fold increase; P = 0.02). Cell proliferation was significantly higher in the PDCs of the PRP group (2.2-fold increase; P < 0.01). In the PDCs, the gene expression levels of Scx, Col1a1, and VEGF were significantly increased by PRP (Scx: 2.0-fold increase, P = 0.01; Col1a1: 5.3-fold increase, P = 0.01; VEGF: 7.8-fold increase, P = 0.01), but the gene expression level of Tnmd, a factor for tendon maturation, was significantly reduced by PRP (0.11-fold decrease; P = 0.02). Conclusion: In vitro PRP promoted migration, proliferation, and tenogenic differentiation with the upregulation of Scx in PDCs. PRP also upregulated the expression of the angiogenic marker VEGF. Clinical Relevance: Our results suggest that PRP treatment in vitro may enhance the tendon-healing properties of PDCs at the initial stage of tendon repair.

Download Full-text

Cytokine Profiling and Intra-Articular Injection of Autologous Platelet-Rich Plasma in Knee Osteoarthritis

International Journal of Molecular Sciences ◽

10.3390/ijms23020890 ◽

2022 ◽

Vol 23 (2) ◽

pp. 890

Author(s):

Kanyakorn Riewruja ◽

Suphattra Phakham ◽

Patlapa Sompolpong ◽

Rangsima Reantragoon ◽

Aree Tanavalee ◽

...

Keyword(s):

Growth Factors ◽

Platelet Rich Plasma ◽

In Vitro Study ◽

Degenerative Joint Disease ◽

Joint Disease ◽

Knee Oa ◽

Oa Chondrocytes ◽

High Level ◽

Cytokine Profiling

Osteoarthritis (OA) is a degenerative joint disease leading to joint pain and stiffness. Due to lack of effective treatments, physical and psychological disabilities caused by OA have a detrimental impact on the patient’s quality of life. Emerging evidence suggests that intra-articular injection of platelet-rich plasma (PRP) may provide favorable results since PRP comprises not only a high level of platelets but also a huge amount of cytokines, chemokines, and growth factors. However, the precise mechanism and standardization method remain uncertain. This study aimed to examine cytokine profiling in both PRP and platelet-poor plasma (PPP) of knee OA patients and to determine the effects of PRP on OA chondrocytes and knee OA patients. PRP contained a wide variety of cytokines, chemokines, growth factors, and autologous intra-articular PRP injection resulted in favorable outcomes in knee OA patients. Significant increases in levels of IL-1, IL-2, IL-7, IL-8, IL-9, IL-12, TNF-α, IL-17, PDGF-BB, bFGF, and MIP-1β were detected in PRP compared to PPP (p < 0.001). An in vitro study showed a marked increase in proliferation in OA chondrocytes cultured with PRP, compared to PPP and fetal bovine serum (p < 0.001). In a clinical study, knee OA patients treated with PRP showed improvement of physical function and pain, assessed by physical performance, Western Ontario and McMaster Universities Arthritis Index and visual analog scale. Our findings from both in vitro and clinical studies suggest that intra-articular PRP injection in knee OA patients may be a potential therapeutic strategy for alleviating knee pain and delaying the need for surgery.

Download Full-text

Comparing the osteogenic potential of schneiderian membrane and dental pulp mesenchymal stem cells: an in vitro study

Cell and Tissue Banking ◽

10.1007/s10561-020-09887-4 ◽

2021 ◽

Author(s):

Antoine Berbéri ◽

Joseph Sabbagh ◽

Rita Bou Assaf ◽

Michella Ghassibe-Sabbagh ◽

Fatima Al-Nemer ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Dental Pulp ◽

In Vitro Study ◽

Vitro Study ◽

Osteogenic Potential ◽

Schneiderian Membrane

Download Full-text

Isolation, Characterization, and Differentiation of Stem Cells From Various Dental Sources: An In Vitro Study

Journal of Advanced Oral Research ◽

10.1177/23202068211010768 ◽

2021 ◽

pp. 232020682110107

Author(s):

Sandeep S. Katti ◽

Kishore Bhat ◽

Chetana Bogar

Keyword(s):

Stem Cells ◽

Growth Factors ◽

Statistical Analysis ◽

Specific Growth ◽

Culture Media ◽

In Vitro Study ◽

Central Research ◽

Cell Lineages ◽

Apical Papilla

Aim: The aim of the current study was to isolate stem cells from various dental sources such as dental pulp, periodontal ligament (PDL), and apical papilla, and to characterize stem cells by staining for the presence/absence of specific surface markers and also to differentiate stem cells into osteogenic, chondrogenic, and adipogenic cell lineages by exposing them to specific growth factors under the ideal conditions. Materials and Methods: A total of 117 samples were included in the study, consisting of 30 pulp, 50 gingival, 35 PDL, and 2 apical papilla samples. The pulp was extirpated and transported to the Central Research Laboratory. Gingival connective tissue was collected from the participants undergoing any crown lengthening procedure or any gingivectomy procedure from the Department of Periodontology. A similar procedure was also followed for apical papilla and PDL. Isolation was done followed by the identification of the cells by immunocytochemistry using different markers. Once the identity of cells was confirmed, these cells were treated with different culture media to attain 70% to 100% confluency. Then the medium was replaced with a conditioning medium containing specific growth factors for differentiation into osteogenic, chondrogenic, and adipogenic cell lineages. Result: In our study, the number of samples collected and processed was 117. The isolation rate of stem cells from the above-collected samples was 70%. Statistical analysis—no statistical analysis was done as there was no variability expected. Conclusion: Our study showed that stem cells could be isolated, differentiated, and characterized from different dental sources.

Download Full-text

Evaluation of coconut water neutralized by different agents on the viability of human fibroblasts: an in vitro study

Revista de Odontologia da UNESP ◽

10.1590/1807-2577.09216 ◽

2016 ◽

Vol 45 (4) ◽

pp. 234-239 ◽

Cited By ~ 1

Author(s):

Priscilla Barbosa Ferreira SOARES ◽

Aletheia Moraes ROCHA ◽

Manuella Verdinelli de Paula REIS ◽

Camilla Christian Gomes MOURA ◽

Carlos José SOARES

Keyword(s):

Cell Viability ◽

Tap Water ◽

Human Fibroblasts ◽

In Vitro Study ◽

Positive Control ◽

Negative Control ◽

Coconut Water ◽

Ph Adjustment ◽

Adjustment Method

Abstract Objective This study evaluated four types of pH adjustment of the coconut water (CW) on viability of human fibroblasts (HFF). Material and method Natural and industrialized CW were adjusted to pH 7.0 using: (1) Sodium Hidroxide (NaOH), (2) Sodium bicarbonate (NaHCO3), (3) Triethanolamine (C6H15NO3), (4) 2-Amino-2-Methil-1-Propanol (C4H11NO). Fibroblasts were plated at 2×104/ well in 96 well plates and maintained in the CW solutions for 2 h and 4 h. Positive control was represented by HFF maintained in DMEM and the negative control by tap water. Cell viability was analyzed by MTT formazan method. Data were analyzed by 3-way ANOVA followed by Tukey’s and Dunnet’s test. Result There are no significant effect on the cell viability regarding type of CW, period of evaluation, and the interactions between CW and period of evaluation, CW and pH adjustment method, pH adjustment method and period of evaluation (p>0.05). Conclusion The product used for CW pH adjustment did not influenced HFF viability, thought there are a tendency of better performance in natural CW.

Download Full-text