Abstract : Spectrophotometric determination of platelet counts in platelet-rich plasma
Platelet-rich plasma (PRP) is widely used in regenerative dentistry and other medical fields. However, its effectiveness has often been questioned. For better evaluation, the quality of individual PRP preparations should be assured prior to use. We proposed a spectrophotometric method for determination of platelet counts and validated its applicability using two types of PRP preparations.
Blood samples were obtained from healthy male volunteers and pure PRP (P-PRP) and leukocytes-rich PRP (L-PRP) were prepared using the double-spin method. In serial dilutions, platelet counts in P-PRP and L-PRP were determined using an automated hematology analyzer and a compact spectrophotometer. For validation, P-PRP and L-PRP independently prepared by three well-trained operators were used for comparison of the calculated and measured platelet counts.
In the two types of PRP samples evaluated, platelet counts were almost equal and greater amount of both white blood cells (WBCs) and red blood cells (RBCs) were included in L-PRP preparations. The calibration curve obtained from serially diluted P-PRP showed a strong correlation (R2 = 0.995), whereas that of L-PRP was relatively weaker (R2 = 0.975). In validation testing, the scatter plot of the calculated platelet counts versus the measured values showed a strong correlation in P-PRP (R2 = 0.671), whereas that of L-PRP showed a much weaker correlation (R2 = 0.0605).
This method can precisely determine platelet counts in PRP preparations when the inclusion of WBCs or RBCs is minimized. Therefore, we recommend that clinicians use this method for quality assurance of individual PRP preparations.
Serial posts:
- Abstract : Spectrophotometric determination of platelet counts in platelet-rich plasma
- Background : Spectrophotometric determination of platelet counts in platelet-rich plasma [1]
- Background : Spectrophotometric determination of platelet counts in platelet-rich plasma [2]
- Methods : Spectrophotometric determination of platelet counts in platelet-rich plasma [1]
- Methods : Spectrophotometric determination of platelet counts in platelet-rich plasma [2]
- Results : Spectrophotometric determination of platelet counts in platelet-rich plasma [1]
- Results : Spectrophotometric determination of platelet counts in platelet-rich plasma [2]
- Discussion : Spectrophotometric determination of platelet counts in platelet-rich plasma [1]
- Discussion : Spectrophotometric determination of platelet counts in platelet-rich plasma [2]
- Discussion : Spectrophotometric determination of platelet counts in platelet-rich plasma [3]
- Conclusions : Spectrophotometric determination of platelet counts in platelet-rich plasma
- Abbreviations : Spectrophotometric determination of platelet counts in platelet-rich plasma
- References : Spectrophotometric determination of platelet counts in platelet-rich plasma [1]
- References : Spectrophotometric determination of platelet counts in platelet-rich plasma [2]
- References : Spectrophotometric determination of platelet counts in platelet-rich plasma [3]
- References : Spectrophotometric determination of platelet counts in platelet-rich plasma [4]
- Availability of data and materials : Spectrophotometric determination of platelet counts in platelet-rich plasma
- Author information : Spectrophotometric determination of platelet counts in platelet-rich plasma [1]
- Author information : Spectrophotometric determination of platelet counts in platelet-rich plasma [2]
- Ethics declarations : Spectrophotometric determination of platelet counts in platelet-rich plasma
- Rights and permissions : Spectrophotometric determination of platelet counts in platelet-rich plasma
- About this article : Spectrophotometric determination of platelet counts in platelet-rich plasma
- Fig. 1. A compact SPM with its remote controller installed on an iPad Air. iPhones and other Android devices can be used instead of the iPad Air : Spectrophotometric determination of platelet count
- Fig. 2. The appearance of blood sampled after gravity fractionation and the resulting P-PRP and L-PRP. In the first low-speed spin, samples were centrifuged for 10 min at 533×g. For P-PRP preparation, the upper plasma fraction, which was 2 mm beyond the interface between plasma and RBC fractions, was transferred into sample tubes for the second high-speed spin (2656×g, 5 min). In contrast, for L-PRP preparation, the upper plasma fraction including the buffy coat and the surface of the RBC fraction was used for the second spin. The supernatant (PPP) was excluded by 50–70%, and platelets were resuspended in the remaining PPP fraction : Spectrophotometric determination of platelet count
- Fig. 3. Counts of platelets (PLT), WBCs, and RBCs in P-PRP and L-PRP preparations prepared for calibration curves. N = 14 for each type of PRP : Spectrophotometric determination of platelet count
- Fig. 4. Calibration curves of measured platelet counts versus absorbance in P-PRP and L-PRP preparations. The samples were serially diluted by PPP, and the platelet counts were determined using an AHA and SPM. N = 14 for each type of PRP : Spectrophotometric determination of platelet count
- Fig. 5. Counts of platelets (PLT), WBCs, and RBCs in P-PRP and L-PRP preparations prepared for validation testing. N = 32 and 50 for P-PRP and L-PRP, respectively : Spectrophotometric determination of platelet count
- Fig. 6. Scatter plots representing possible correlations between platelet (PLT) and WBC counts and between platelet and RBC counts in P-PRP and L-PRP preparations. Note: strong positive correlations were observed between platelets and RBC in both PRP types. N = 32 and 50 for P-PRP and L-PRP, respectively : Spectrophotometric determination of platelet count
- Fig. 7. Scatter plots representing correlations between measured and calculated platelet counts in P-PRP and L-PRP preparations. Note: a strong correlation was observed only in P-PRP. N = 32 and 50 for P-PRP and L-PRP, respectively : Spectrophotometric determination of platelet count