Discussion : Spectrophotometric determination of platelet counts in platelet-rich plasma [2]
Another limitation is the color of plasma. In terms of color, blood samples obtained from the donors participating in this study were light yellow and could be evaluated as “normal.” However, we have sometimes encountered colored plasma samples in clinical practice. For example, when blood triglyceride levels are high, the plasma turns milky white or turbid [22,23,24]. Hemolytic plasma looks reddish, while icteric plasma appears yellow. When the degree of color change is not severe and when the transparency is maintained, the data may be compensated by the absorbance of PPP. However, in this case, we recommend the use of an AHA for accurate determination of the platelet counts.
We should discuss briefly how clinicians can perform quality assurance for individual PRP preparations. As described elsewhere [10, 25], PRP quality is evaluated mainly based on two major points: sterility and efficacy. Recent advances in PCR technology enable clinicians to quickly assess the contamination of targeted bacteria and mycoplasmas [26] in clinical settings. However, clinicians may require a well-trained operator for this kind of sterility testing. The current regulatory framework for PRP therapy in Japan requires clinicians to prepare PRP on a clean bench [10]. Therefore, as long as blood samples are handled aseptically, the resulting PRP preparations are evaluated as sterile.
As for efficacy, regardless of the assay system, several hours or days are required to complete efficacy testing. Even if it takes only several hours, unfortunately, this delay is not beneficial to many patients and is not suitable for on-site preparation and immediate use in autologous PRP therapy. The only exception is platelet counting, which takes only a few minutes with the use of an AHA. However, it is a problem that the conventional form of this device is $10,000 or higher and requires installation space (500 × 500 mm at least). In contrast, the compact SPM used in this study costs only $800 and can be stored in a drawer. Therefore, despite several limitations, this compact SPM would be useful for fundamental quality assurance as well as for the examination of possible correlations between platelet counts and clinical outcomes.
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
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- 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